Photoshop_for_ResearchPSHOP FOR RESEARCH - READMETEXTstopStopStopStopMsgeTEXTAll Actions work with images that are already open, EXCEPT for Multi-Image Layout actions, which require that the screen is cleared so that images can be opened from directories.CntnboolTEXTstopStopStopStopMsgeTEXTXForinfo on each set of actions, click buttons that contain all capitalized lettering...CntnboolTEXTstopStopStopStopMsgeTEXTSome actions also contain "+ info" in the title as a way to inform the user. A second action is often made sans the info to reduce interactivity.CntnboolTEXTstopStopStopStopMsgeTEXTIt is assumed that these actions will be used in button mode. If these are not in button mode, then click on the upper, right arrowhead of the Actions dialogue box and choose "Button Mode" from the drop down list.TROUBLESHOOTING INFOTEXTstopStopStopStopMsgeTEXTWhen these actions stop suddenly with what is usually a prompt unrelated to the problem, figure that one of 3 things are wrong: Mode is in indexed color when it should be RGB color or Grayscale...CntnboolTEXTstopStopStopStopMsgeTEXTThe mode is 16- or 48-bit when it should be 8- or 24-bit. Or the image is layered as a Photoshop file and the action is running on the wrong layer, such as a text or shape layer...CntnboolTEXTstopStopStopStopMsgeTEXTAll actions require the use of non-layered files. To eliminate layers, even if these do not show in the Layers dialogue box (Window > Layers), use the Flatten All Layers action, or Layer > Flatten Image...CntnboolTEXTstopStopStopStopMsgeTEXTIf there is text or shapes (such as arrowheads) on your image and you wish to preserve the layers for future editing, then save the "editable" image file as a layered, Photoshop formatted file and flatten a copy for use with these actions.preferences-disable inquiryTEXTsetSetSetsetdnullobj propPrpr colorSettingsEnmrcappOrdnTrgtT Objc colorSettings workingRGBTEXTAdobe RGB (1998)askMismatchOpeningboolTEXTsetSetSetsetdnullobj propPrprFlSPEnmrcappOrdnTrgtT ObjcFlSvExtQenumQurSQurATEXTsetSetSetsetdnullobj propPrprTrnPEnmrcappOrdnTrgtT ObjcTrnPTrnGenumTrnGNoneTEXTstopStopStopStopMsgeTEXTThis action only needs to be run once. It sets the Color Settings box to Adobe 1998 color settings and turns off prompts in regard to mismatched color settings. It also turns off the grid pattern to show transparency, something confusing to many users.CntnboolOPEN FILE - CHANGE MODE + INFOTEXTstopStopStopStopMsgeTEXTActions that follow provide interactive open function so that your image is duplicated (to prevent saving over existing file). Also, changes in the type or mode of image from 16-bit to 8-bit and to RGB Color. F2 Open - Duplicate ImageTEXTopenOpenOpenOpn nullPth ]6D:\#Personal_files\science_images\h&e-dab\Dab_20xb.tif;Pf ób0+- -\f ,Pf C2C ]#2RC#2CCx]]<]wACTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls  ModeGrayscale16- to 8-bit +infoTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtInptVlLslonglongTEXT convertMode Convert Mode Convert ModeCnvMDpthlongTEXTstopStopStopStopMsgeTEXTTYour image still may not be bright enough. Use actions later on to brighten image.Mode Grayscale 16- to 8-bitTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtInptVlLslonglongTEXT convertMode Convert Mode Convert ModeCnvMDpthlongMode RGB ColorTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMMode Indexed ColorTEXT convertMode Convert Mode Convert ModeCnvMT ObjcIndCPlt enumClrPAdptClrslongDthqenumDthqDthbCOLOR TO GRAYSCALE + INFOTEXTstopStopStopStopMsgeTEXTThese actions take a color image and return it to grayscale. If your image is colorized, be sure to choose from red, green or blue to grayscale so that luminance or its perceived brightness is not sacrificed!CntnboolTEXTstopStopStopStopMsgeTEXTYou can also split a color image into its respective grayscale channels by choosing "Split Channels." The best grayscale image can be chosen from the red, green or blue grayscale channel.CntnboolTEXTstopStopStopStopMsgeTEXTIf you use "RGB Color to Grayscale" then Photoshop will determine best grayscale rendition of the color image. This works well for brightfield images (though split channels should be tried), but not for colorized, darkfield images. From_Red_toGrayTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxRd UntF#Prc@YFrom_Green_toGrayTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxGrn UntF#Prc@YFrom_Blue_toGrayTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxBl UntF#Prc@YSplit ChannelsTEXT splitChannelsSplit ChannelsRGB Color to GrayscaleTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysROTATE & FLIP IMAGE + INFOTEXTstopStopStopStopMsgeTEXTvThe following methods allow you to rotate the entire image in theta, or to flip the image horizontally or vertically.Arbitrary RotationTEXTrotateEventEnumRotateRotateRttenullobj EnmrDcmnOrdnFrstAnglUntF#Ang@ Rotate 180TEXTrotateEventEnumRotateRotateRttenullobj EnmrDcmnOrdnFrstAnglUntF#Ang@f Rotate 90 CWTEXTrotateEventEnumRotateRotateRttenullobj EnmrDcmnOrdnFrstAnglUntF#Ang@VRotate 90 CCWTEXTrotateEventEnumRotateRotateRttenullobj EnmrDcmnOrdnFrstAnglUntF#AngVFlip VerticalTEXTflipFlipFlipFlipnullobj EnmrDcmnOrdnFrstAxisenumOrntVrtcFlip HorizontalTEXTflipFlipFlipFlipnullobj EnmrDcmnOrdnFrstAxisenumOrntHrznFILTERS - BLUR SHARPEN VIDEOTEXTstopStopStopStopMsgeTEXTThe following actions can be used to sharpen, blur, remove dust, or deinterlace analog video. Except for the deinterlace action, apply these more than once to increase filtering.Sharpen lowestTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMRds doub?Amntlong2Thshlong Sharpen_lowTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMRds doub?AmntlongKThshlongSharpen mediumTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMRds doub?AmntlongThshlong Sharpen highTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMRds doub?Amntlong,ThshlongSharpen highestTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMRds doub?AmntlongThshlongSharpen InteractiveTEXT unsharpMask Unsharp Mask Unsharp MaskUnsMAmntUntF#Prc@RRds UntF#Pxl?ThshlongMedian (blur) interactiveTEXTmedianMedianMedianMdn Rds UntF#Pxl?Video - DeinterlaceTEXTstopStopStopStopMsgeTEXTUse this filter if you are using an ANALOG video cable to your computer. Try this filter, then zoom in to determine whether it has improved the image. Check "before and after" by using the Edit > Undo Redo or by using cntrl/cmd key + Z. CntnboolTEXT deInterlace De-Interlace De-InterlaceDntrIntEenumIntEElmOIntCenumIntCCrtIBACKGROUND CORRECTION + INFOTEXTstopStopStopStopMsgeTEXTThese actions remove background artifacts or even out the illumination (flatfield or shading correction) from images taken through microscopes, EM,gels, etc. Remove Dust 1TEXTdustAndScratchesDust & ScratchesDust & ScratchesDstSRds longThshlongRemove Dust 2TEXTdustAndScratchesDust & ScratchesDust & ScratchesDstSRds longThshlongPhase DIC background cleanerTEXThighPass High Pass High PassHghPRds UntF#Pxl@D&f`Background cleaner interactiveTEXThighPass High Pass High PassHghPRds UntF#Pxl@D&f`Gel background flatteningTEXThighPass High Pass High PassHghPRds UntF#Pxl@D&f`shading correction + infoTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTstopStopStopStopMsgeTEXTFirst you will get a "Gaussian Blur" dialogue box. Set the blurring with the slider so that details cannot be made out within features, but not so much that the blur creates a border that extends much beyond the original edge.CntnboolTEXT gaussianBlur Gaussian BlurGaussian BlurGsnBRds UntF#Pxl@%TEXTinvertInvertTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMHrdLTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@2TEXTstopStopStopStopMsgeTEXTUse the Opacity slider in the Layer's Adjustment dialogue box to set value. Iteratively change the value until it appears that the background is evenly illuminated. CntnboolTEXTstopStopStopStopMsgeTEXT Move cursor over background and check values in the Info box to see where the Opacity Slider should be set. Values in the background should be nearly similar for the K: value or one of the 3 RGB values no matter where the cursor is placed.CntnboolTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@BTEXT flattenImage Flatten ImageTEXTstopStopStopStopMsgeTEXTNow adjust white levels, gamma and black levels in the Levels dialogue box. If the image needs to be color corrected, use the white eyedropper tool and click in white area (or what should be white) of background.CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtOtptVlLslonglongshading correction TEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXT gaussianBlur Gaussian BlurGaussian BlurGsnBRds UntF#Pxl@%TEXTinvertInvertTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMHrdLTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@2TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@BTEXT flattenImage Flatten ImageTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtOtptVlLslonglongCOLOR CONTRAST BRIGHTFIELDTEXTstopStopStopStopMsgeTEXTThe 1st series of autobalance actions increase the black level to varying degrees so that shadow areas will reproduce for various outputs. The brightness and color are autobalanced in each: only the black level differs by degrees.CntnboolTEXTstopStopStopStopMsgeTEXTThe black level is often difficult to reproduce, thus the varying levels of black given as choices. You should check the black level in your darkest areas...CntnboolTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTInfo Box K: values in the darkest areas should read at 85 - 90% and RGB values at 20 - 35 (R, G, B values may differ if image is in color: 20 would be the LOWEST value for any one channel).CntnboolTEXTstopStopStopStopMsgeTEXTXThe interactive actions contain prompts when the "+ info" is included with the title. CntnboolTEXTstopStopStopStopMsgeTEXTvThe term "brightfield" is meant to include the kinds of images in which features are dark against a light background.Contrast/color-autobalance 1TEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsOtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlRd AutoboolObjcLvlAChnlobj EnmrChnlChnlGrn AutoboolObjcLvlAChnlobj EnmrChnlChnlBl AutoboolContrast/color-autobalance 2TEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsOtptVlLslong longObjcLvlAChnlobj EnmrChnlChnlRd AutoboolObjcLvlAChnlobj EnmrChnlChnlGrn AutoboolObjcLvlAChnlobj EnmrChnlChnlBl AutoboolContrast/color-autobalance 3TEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsOtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlRd AutoboolObjcLvlAChnlobj EnmrChnlChnlGrn AutoboolObjcLvlAChnlobj EnmrChnlChnlBl AutoboolContrast/color-autobalance 4TEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsOtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlRd AutoboolObjcLvlAChnlobj EnmrChnlChnlGrn AutoboolObjcLvlAChnlobj EnmrChnlChnlBl AutoboolContrast/color-autobalance 5TEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsOtptVlLslong(longObjcLvlAChnlobj EnmrChnlChnlRd AutoboolObjcLvlAChnlobj EnmrChnlChnlGrn AutoboolObjcLvlAChnlobj EnmrChnlChnlBl AutoboolIncrease Shadow Detail + infoTEXTstopStopStopStopMsgeTEXTApply this action more than once to lighten shadows. This action, unlike contrast/color autobalance actions, does not correct for color: it affects gamma (thereby affecting mostly midtones) to lighten shadows.CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?񙙙OtptVlLslonglongIncrease Shadow DetailTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?񙙙OtptVlLslonglongSelect Colors to Adjust + InfoTEXTstopStopStopStopMsgeTEXTqColor Range allows you to select certain hues or gray levels, thus limiting the range of colors/grays to alter. CntnboolTEXTstopStopStopStopMsgeTEXTThe best results are had by first clicking on an area of your image containing the hue/gray level of interest. Then choose the "plus" eyedropper tool to click on several (the more the better) places on your image with similar hues or gray levels.CntnboolTEXTstopStopStopStopMsgeTEXTThe fuzziness slider can be increased to include more hues or gray levels, but it will also introduce fuzzy edges (a concern when subsequently measuring the selected area).CntnboolTEXTstopStopStopStopMsgeTEXTIf you unintentionally click on a hue or gray level on your image and select too much (you will see it on the preview window), then it may be best to start over again. CntnboolTEXTstopStopStopStopMsgeTEXTFYou can save what you have selected to apply to other similar images.CntnboolTEXT colorRange Color Range Color RangeClrRFznslong Mnm ObjcLbClLmncdoub@X =pA doubB doubMxm ObjcLbClLmncdoub@X =pA doubB doubSelect Colors to AdjustTEXT colorRange Color Range Color RangeClrRFznslong Mnm ObjcLbClLmncdoub@X =pA doubB doubMxm ObjcLbClLmncdoub@X =pA doubB doubContrast Color adjust + infoTEXTstopStopStopStopMsgeTEXTIf image is in color, choose the white eyedropper tool and click on a white area of the image to auto balance colors. This may have to be done iteratively while choosing different areas in your image until the colors look correct.CntnboolTEXTstopStopStopStopMsgeTEXTIf your background is uneven, color correction may be difficult to apply. Use the shading correction action earlier in this set.CntnboolTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTiInfo Box K: values in the darkest areas should read at 85 - 90% and minimum R, G, or B value at 20 - 35.CntnboolTEXTstopStopStopStopMsgeTEXTTo LIGHTEN black levels, use the black OUTPUT slider and move to the right, iteratively checking black level on image and looking at Info box. To DARKEN black levels, use the black INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXTLimit the top end of the white values in your image by keeping white OUTPUT slider at 254. You may increase white values by moving the white INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXT$Adjust gamma INPUT slider by eye. CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?\(Contrast Color adjustTEXTstopStopStopStopMsgeTEXTIf image is in color, choose the white eyedropper tool and click on a white area of the image to auto balance colors. This may have to be done iteratively while choosing different areas in your image until the colors look correct.CntnboolTEXTstopStopStopStopMsgeTEXTIf your background is uneven, color correction may be difficult to apply. Use the shading correction action earlier in this set.CntnboolTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTInfo Box K: values in the darkest areas should read at 85 - 90% and RGB values at 20 - 35 (R, G, B values may differ if image is in color: 20 would be the HIGHEST value for any one channel).CntnboolTEXTstopStopStopStopMsgeTEXTTo LIGHTEN black levels, use the black OUTPUT slider and move to the right, iteratively checking black level on image and looking at Info box. To DARKEN black levels, use the black INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXTLimit the top end of the white values in your image by keeping white OUTPUT slider at 254. You may increase white values by moving the white INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXT$Adjust gamma INPUT slider by eye. CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?\(Contrast-adjust curvesTEXTcurvesCurvesCurvesCrvsColor-variationsTEXT variations Variations VariationsVrtn RdGmdoub?GrnGdoub?BlGmdoub?RdWhlongGrnWlongBlWhlongRdBllongGrnBlongBlBllongStrtlongdColor-hue & saturation + infoTEXTstopStopStopStopMsgeTEXTThe most common problem with color in brightfield images from light microscopes lies with the magenta tones. Often these are far too "neon" or saturated. Thus, in Hue & Saturation, the Saturation slider is normally decreased with the slider.CntnboolTEXTstopStopStopStopMsgeTEXTChoose the color you wish to change from the drop down list, and then expand, if necessary, using sliders at the bottom (drag from middle bar of sliders).CntnboolTEXT hueSaturationHue/SaturationHue/SaturationHStrClrzboolColor-hue & saturationTEXT hueSaturationHue/SaturationHue/SaturationHStrClrzboolBlue- correct color + infoTEXTstopStopStopStopMsgeTEXTThis action corrects for blue- to blue-violet colors, normally irreproducable on output devices. The colors are shifted to more of a "printable" cyan blue.CntnboolTEXT replaceColor Replace ColorReplace ColorRplCFznslongMnm ObjcLbClLmncdoub@'QA doub@8B doubHMxm ObjcLbClLmncdoub@'QA doub@8B doubHH longStrtlongLghtlongBlue- correct colorTEXT replaceColor Replace ColorReplace ColorRplCFznslongMnm ObjcLbClLmncdoub@'QA doub@8B doubHMxm ObjcLbClLmncdoub@'QA doub@8B doubHH longStrtlongLghtlongPSEUDOCOLOR + INFOTEXTstopStopStopStopMsgeTEXThPsuedocolor is often used to draw attention to optical varying levels of optical densities in an image.!Pseudocolor-spectrum + info copyTEXTstopStopStopStopMsgeTEXTThis action must first turn your image into grayscale. If your image is in color, it is best that you convert to grayscale yourself by using Channel Mixer, or by using "Make Grayscale from Red (Green or Blue)" actions.CntnboolTEXTstopStopStopStopMsgeTEXTPsuedocolor spectrum uses the colors from the visible spectrum to colorize a grayscale image, with colors assigned according to optical density values of image. The darkest are assigned violet to blue, the brightest red.CntnboolTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXT convertMode Convert Mode Convert ModeCnvMT typeIndCTEXTsetSetSetsetdnullobj propClr ClrTT enumClrPSpctTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT duplicate Duplicate DuplicateDplcnullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj propLyr BckgTEXTpastePasteTEXThideHideHideHd nullVlLsobj nameLyr Background copyTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr Background copyTEXTselectSelectSelectslctnullobj nameLyr Background copyPseudocolor-spectrum copy TEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXT convertMode Convert Mode Convert ModeCnvMT typeIndCTEXTsetSetSetsetdnullobj propClr ClrTT enumClrPSpctTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT duplicate Duplicate DuplicateDplcnullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj propLyr BckgTEXTpastePasteTEXThideHideHideHd nullVlLsobj nameLyr Background copyTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr Background copyTEXTselectSelectSelectslctnullobj nameLyr Background copyPseudocolor-8 grays copy TEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXT posterization Posterize PosterizePstrLvlslongTEXT duplicate Duplicate DuplicateDplcnullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj propLyr BckgTEXTpastePasteTEXThideHideHideHd nullVlLsobj nameLyr Background copyTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr Background copyTEXTselectSelectSelectslctnullobj nameLyr Background copyPseudocolor-8 colors copy TEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXT posterization Posterize PosterizePstrLvlslongTEXT duplicate Duplicate DuplicateDplcnullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj propLyr BckgTEXTpastePasteTEXThideHideHideHd nullVlLsobj nameLyr Background copyTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr Background copyTEXTselectSelectSelectslctnullobj nameLyr Background copy COLOR CONTRAST DARKFIELD + INFOTEXTstopStopStopStopMsgeTEXTJDarkfield images include images of bright objects on a dark background. CntnboolTEXTstopStopStopStopMsgeTEXTlThe black level is often difficult to reproduce. You should check the black level in your darkest areas...CntnboolTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTInfo Box K: values in the darkest areas should read at 85 - 90% and RGB values at 20 - 35 (R, G, B values may differ if image is in color: 20 would be the LOWEST value for any one channel).CntnboolTEXTstopStopStopStopMsgeTEXTXThe interactive actions contain prompts when the "+ info" is included with the title. CntnboolFluorescence-brighter + infoTEXTstopStopStopStopMsgeTEXT>Click more than once to increase brightness to desired value.CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtInptVlLslonglongOtptVlLslonglongFluorescence-brighterTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlOrdnTrgtInptVlLslonglongOtptVlLslonglongFluorescence- up shadow + infoTEXTstopStopStopStopMsgeTEXTThis protocol uses Curves to brighten shadow and background detail while also increasing contrast in whiter midtones. Apply more than once to brighten more. CntnboolTEXTstopStopStopStopMsgeTEXTAs you increase brightness, be sure to move cursor over background areas to see that the K: value in the Info box reads 85 - 90% and/or the minimum R, G, or B value reads 20 - 35.CntnboolTEXTcurvesCurvesCurvesCrvsAdjsVlLsObjcCrvAChnlobj EnmrChnlOrdnTrgtCrv VlLsObjcPnt HrzndoubVrtcdoubObjcPnt Hrzndoub@JVrtcdoub@PObjcPnt Hrzndoub@h Vrtcdoub@fObjcPnt Hrzndoub@oVrtcdoub@oFluorescence- up shadowTEXTcurvesCurvesCurvesCrvsAdjsVlLsObjcCrvAChnlobj EnmrChnlOrdnTrgtCrv VlLsObjcPnt HrzndoubVrtcdoubObjcPnt Hrzndoub@JVrtcdoub@PObjcPnt Hrzndoub@h Vrtcdoub@fObjcPnt Hrzndoub@oVrtcdoub@oFluorescence- up contrastTEXTcurvesCurvesCurvesCrvsAdjsVlLsObjcCrvAChnlobj EnmrChnlChnlCmpsCrv VlLsObjcPnt HrzndoubVrtcdoubObjcPnt Hrzndoub@XVrtcdoub@TObjcPnt Hrzndoub@e@Vrtcdoub@gObjcPnt Hrzndoub@oVrtcdoub@oBrighten Shadow Detail + infoTEXTstopStopStopStopMsgeTEXTApply this action more than once to lighten shadows. This action affects gamma (thereby affecting mostly midtones) to lighten shadows. It also uses Levels versus Curves and does not simultaneously increase contrast in white areas.CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?񙙙OtptVlLslonglongBrighten Shadow DetailTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?񙙙OtptVlLslonglongSelect Colors to Adjust + InfoTEXTstopStopStopStopMsgeTEXTqColor Range allows you to select certain hues or gray levels, thus limiting the range of colors/grays to alter. CntnboolTEXTstopStopStopStopMsgeTEXTThe best results are had by first clicking on an area of your image containing the hue/gray level of interest. Then choose the "plus" eyedropper tool to click on several (the more the better) places on your image with similar hues or gray levels.CntnboolTEXTstopStopStopStopMsgeTEXTThe fuzziness slider can be increased to include more hues or gray levels, but it will also introduce fuzzy edges (a concern when subsequently measuring the selected area).CntnboolTEXTstopStopStopStopMsgeTEXTIf you unintentionally click on a hue or gray level on your image and select too much (you will see it on the preview window), then it may be best to start over again. CntnboolTEXTstopStopStopStopMsgeTEXTFYou can save what you have selected to apply to other similar images.CntnboolTEXT colorRange Color Range Color RangeClrRFznslong Mnm ObjcLbClLmncdoub@X =pA doubB doubMxm ObjcLbClLmncdoub@X =pA doubB doubSelect Colors to AdjustTEXT colorRange Color Range Color RangeClrRFznslong Mnm ObjcLbClLmncdoub@X =pA doubB doubMxm ObjcLbClLmncdoub@X =pA doubB doub Contrast adjust manually + infoTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTiInfo Box K: values in the darkest areas should read at 85 - 90% and minimum R, G, or B value at 20 - 35.CntnboolTEXTstopStopStopStopMsgeTEXTTo LIGHTEN black levels, use the black OUTPUT slider and move to the right, iteratively checking black level on image and looking at Info box. To DARKEN black levels, use the black INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXTLimit the top end of the white values in your image by keeping white OUTPUT slider at 254. You may increase white values by moving the white INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXT$Adjust gamma INPUT slider by eye. CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?\(Contrast adjust manuallyTEXTstopStopStopStopMsgeTEXTIf image is in color, choose the white eyedropper tool and click on a white area of the image to auto balance colors. This may have to be done iteratively while choosing different areas in your image until the colors look correct.CntnboolTEXTstopStopStopStopMsgeTEXTIf your background is uneven, color correction may be difficult to apply. Use the shading correction action earlier in this set.CntnboolTEXTstopStopStopStopMsgeTEXTTo check the level of the darkest areas on your image, place cursor over darkest area and read from the info box (be sure its "Palette Options" have been set to 1st Readout: Grayscale, 2nd Readout: RGB Color).CntnboolTEXTstopStopStopStopMsgeTEXTInfo Box K: values in the darkest areas should read at 85 - 90% and RGB values at 20 - 35 (R, G, B values may differ if image is in color: 20 would be the HIGHEST value for any one channel).CntnboolTEXTstopStopStopStopMsgeTEXTTo LIGHTEN black levels, use the black OUTPUT slider and move to the right, iteratively checking black level on image and looking at Info box. To DARKEN black levels, use the black INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXTLimit the top end of the white values in your image by keeping white OUTPUT slider at 254. You may increase white values by moving the white INPUT slider.CntnboolTEXTstopStopStopStopMsgeTEXT$Adjust gamma INPUT slider by eye. CntnboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlCmpsGmm doub?\( Contrast adjust manually CurvesTEXTcurvesCurvesCurvesCrvsDE-COLORIZE DARKFIELD IMAGESTEXTstopStopStopStopMsgeTEXTThese actions de-colorize (return to grayscale) images that have been saved as colorized images from confocal software. These colorized images are often saturated. Thus, these need to be colorized again with less saturated colors.CntnboolTEXTstopStopStopStopMsgeTEXTColorizing with colors that reproduce well for various outputs can be found in the actions in the COLORIZE section following this.TEXTstopStopStopStopMsgeTEXTPDarkfield images include those that have lighter features on a dark background. De-Colorize Red (MakeGrayscale)TEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxRd UntF#Prc@YDe-Colorize GreenTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxGrn UntF#Prc@YDe-Colorize BlueTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT channelMixer Channel MixerChannel MixerChnMMnchboolGry ObjcChMxBl UntF#Prc@YCOLORIZING-MERGING DARKFIELDTEXTstopStopStopStopMsgeTEXTImages that are colorized in confocal software often contain values that do not reproduce well, especially in the blue range...CntnboolTEXTstopStopStopStopMsgeTEXTThe following set of colorizing actions, except for the actions marked "saturate," use colors that generally reproduce well...CntnboolTEXTstopStopStopStopMsgeTEXTIf, however, the original image was collected with white levels that read at 255 pixel value (saturated whites) over broad areas of the image, then don't expect good reproduction...CntnboolTEXTstopStopStopStopMsgeTEXTCare must be taken when collecting to avoid saturation in the white or black end of the confocal's dynamic range, except for small features (such as bacteria).Colorize Green (shifted bluer)TEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMFlttboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlRd OtptVlLslonglongnObjcLvlAChnlobj EnmrChnlChnlBl OtptVlLslonglong Colorize Green (shifted yellow)TEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMFlttboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlRd OtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlBl OtptVlLslonglong Colorize Green (saturate) +infoTEXTstopStopStopStopMsgeTEXTA saturated green looks good on a computer screen but typically does not reproduce well for various outputs. This is the same colorized green provided by most confocal software applications.CntnboolTEXTstopStopStopStopMsgeTEXTBoth the previous actions in which colors are shifted either bluer or yellow are better choices: both actions yield an image which has a far greater likelihood of reproducing with luminescence equal to what is seen on the screen.CntnboolTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMFlttboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlRd OtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlBl OtptVlLslonglongColorize Green (saturate)TEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMFlttboolTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlRd OtptVlLslonglongObjcLvlAChnlobj EnmrChnlChnlBl OtptVlLslonglong Colorize Blue (shifted to cyan)TEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXTlevelsLevelsLevelsLvlsAdjsVlLsObjcLvlAChnlobj EnmrChnlChnlRd OtptVlLslonglong/$ȲA >/=v=\dz$p=$$z=(=)ҀTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePasteTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghnLefxObjcLefxTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@YTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstMerge red & green TEXTopenOpenOpenOpn nullPth AD:\photoshp4research\manual\quick_photoshop\merg-fibers-green.tifO7 ČTG8 8 ȲA V|o^g <</$ȲA >/=v=\dz$p=$$z=(=)ҀTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePasteTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghnLefxObjcLefxTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr OpctUntF#Prc@YTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstMerge Red, Green & Blue + infoTEXTstopStopStopStopMsgeTEXTThis action assumes that the screen is empty. You will be prompted to open already red-colorized, green-colorized and blue-colorized images. CntnboolTEXTstopStopStopStopMsgeTEXTOpen the red-colorized image first, as the green- colorized image appears more luminous and may need to be minimized in a subsequent step. CntnboolTEXTstopStopStopStopMsgeTEXTDecrease opacity of the green layer, if necessary, to 85 -90% by clicking on the green layer of the merged image and sliding the opacity slider.CntnboolTEXTstopStopStopStopMsgeTEXTWhen the action finishes, you may find it necessary to adjust brightness/contrast with Levels or Curves of individual layers. When satisfied, it is suggested that the file be saved with the "Save As psd, TIFF & JPG" action.CntnboolTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes1.tif2b0+- -\f ,Pf <2< ]R<<<x]]<]wA<TEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes2.tif2b0+-HH -\f H,Pf {2H{ H]R{{{] ]]wA{TEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghnTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes3.tif2b0+-hh -\f h,Pf X2hX h]RXXX] ]]wAXTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghnMerge Red, Green & BlueTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes1.tif2b0+- -\f ,Pf <2< ]R<<<x]]<]wA<TEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes2.tif2b0+-HH -\f H,Pf {2H{ H]R{{{] ]]wA{TEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghnTEXTopenOpenOpenOpn nullPth ]CD:\#Personal_files\photoshop_instruction\Teacher-Photos\Testes3.tif2b0+-hh -\f h,Pf X2hX h]RXXX] ]]wAXTEXT convertMode Convert Mode Convert ModeCnvMT typeGrysTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTcutCutTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Md enumBlnMLghn DELETE EXCESS IMAGE AREA (CROP) Crop + infoTEXTstopStopStopStopMsgeTEXT[Draw selection around the area you wish to preserve (thus removing everything else) first.CntnboolTEXTcropCropCropTEXTcropCropSelection - save + infoTEXTstopStopStopStopMsgeTEXTDraw selection around area of LARGEST feature from a series of companion images (if you are wishing to match dimensions of several images). If you have NOT drawn this already, then click stop button and draw it first. CntnboolTEXTstopStopStopStopMsgeTEXTYou will save this selection, then crop with the action above, then use the next action to load this selection to a companion image. CntnboolTEXTstopStopStopStopMsgeTEXTIn the companion image, move the selection into place around the feature of interest. The selection can be moved by simply placing cursor inside the scrolling box and dragging when marquee tool is selected. CntnboolTEXTstopStopStopStopMsgeTEXTKEEP THIS IMAGE OPEN AS YOU WORK ON COMPANION FILES. The cropping selection is "borrowed" from this image for companion files.CntnboolTEXT duplicate Duplicate DuplicateDplcnullobj propChnlfselNm TEXTcropping selectionSelection - saveTEXT duplicate Duplicate DuplicateDplcnullobj propChnlfselNm TEXTcropping selectionSelection - LoadTEXTsetSetSetsetdnullobj propChnlfselT obj nameChnlcropping selectionnameDcmnpurkinjegreen copyADD SCALE BAR + INFOTEXTstopStopStopStopMsgeTEXTJThe scale bar is best added at this point, if not already on the image...CntnboolTEXTstopStopStopStopMsgeTEXT`The image must be at its original resolution with no pixels subsequently added or subtracted. White Scale BarTEXTstopStopStopStopMsgeTEXTcThis scale bar is NOT calibrated to your image! It is simply a bar with a length you can specify.CntnboolTEXTstopStopStopStopMsgeTEXTYou will need to know the pixel to micron (or other unit) calibration, something you can obtain by imaging a calibration slide with known values and then determining pixel to micron (or other unit) relationship.CntnboolTEXTstopStopStopStopMsgeTEXTYou will first be prompted to make a New file. Set the length of the New file in pixels to match your calibration. E.g., if 50 pixels = 100microns, enter 50 pixels. Do not change the width. that can be changed later. CntnboolTEXTmakeMakeMakeMk Nw ObjcDcmnNm TEXT white barMd typeGrysWdthUntF#Rlt@3333333HghtUntF#Rlt? =p RsltUntF#Rsl@rFl enumFl Wht TEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTfillFillFillFl UsngenumFlCnWht OpctUntF#Prc@YMd enumBlnMNrmlTEXT copyEventCopyTEXTselectSelectSelectslctnullobj releDcmnTEXTpastePastePastepastAntAenumAnntAnnoTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTresetResetResetRsetnullobj propClr ClrsTEXTexchangeExchange ExchangeExchnullobj propClr ClrsTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTstopStopStopStopMsgeTEXTThe next step allows you to transform or alter the width of the scale bar. Click on center box and drag to make smaller or larger. You will probably have to zoom in to make the bar larger.CntnboolTEXTstopStopStopStopMsgeTEXTWhen finished adjusting width, click on check icon at top, right of menu; or click on the marquee tool and then click "apply."CntnboolTEXT transform Transform TransformTrnfnullobj EnmrLyr OrdnTrgtFTcsenumQCStQcsaOfstObjcOffsetOfstHrznUntF#PxlVrtcUntF#Pxl?'΂HghtUntF#Prc@TH qTEXTstopStopStopStopMsgeTEXTNext, type in text for the unit of measurement. Highlight the portion of text you wish to change and type in new text. Move the position of the text by repositioning cursor away from text until cursor changes to"move" icon, then click and drag.CntnboolTEXTmakeMakeMakeMk nullobj ClssTxLrUsngObjcTxLr Txt TEXT25mmwarpObjcwarp warpStyleenum warpStylewarpNone warpValuedoubwarpPerspectivedoubwarpPerspectiveOtherdoub warpRotateenumOrntHrznTxtCObjcPnt HrznUntF#Prc@E GVrtcUntF#Prc@H5 yC textGriddingenum textGriddingNoneOrntenumOrntHrznAntAenumAnntAnCr textShapeVlLsObjc textShapeTEXTenumTEXTPnt TxttVlLsObjcTxttFromlongT longTxtSObjcTxtS$fontPostScriptNameTEXT Arial-BoldMTFntNTEXTArialFntSTEXTBoldScrplongFntTlongSz UntFC@4 syntheticBoldboolsyntheticItalicbool autoLeadingboolHrzSlongdVrtSlongdTrcklongAtKrboolBslnUntFCfontCapsenumfontCapsNrmlbaselineenumbaselineNrmlUndlbool strikethroughboolligatureboololdStyleboolproportionalNumbersboolRtt boolbaselineDirectionenumbaselineDirection withStreammojiZumedoub gridAlignmentenum gridAlignmentromanhyphenationLanguageenumhyphenationLanguageenglishLanguage wariChuCountlong wariChuScaledoub? wariChuWidowlong wariChuOrphanlongnoBreakboolClr ObjcRGBCRd doub@oGrn doub@oBl doub@oFl boolStrkbool fillFirstboolverticalUnderlinePositionenumverticalUnderlinePositionverticalUnderlineRightparagraphStyleRangeVlLsObjcparagraphStyleRangeFromlongT longparagraphStyleObjcparagraphStyleAlgnenumAlg LeftfirstLineIndentUntFC startIndentUntFC endIndentUntFC spaceBeforeUntFC spaceAfterUntFC hyphenateboolhyphenateWordSizelonghyphenatePreLengthlonghyphenatePostLengthlonghyphenateLimitlonghyphenationZonedoub@BhyphenateCapitalizedbooljustificationWordMinimumdoub?陙justificationWordDesireddoub?justificationWordMaximumdoub?G justificationLetterMinimumdoubjustificationLetterDesireddoubjustificationLetterMaximumdoubjustificationGlyphMinimumdoub?justificationGlyphDesireddoub?justificationGlyphMaximumdoub? hangingRomanbool burasagariboolpreferredKinsokuOrderenumpreferredKinsokuOrderpushIn mojiKumiNameTEXTNonetextEveryLineComposerboolautoLeadingPercentagedoub?33@ leadingTypeenum leadingType leadingBelowTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 1TEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTstopStopStopStopMsgeTEXTMove the position of the scale bar and text above it. These layers are linked, so if you move the scale bar the text will move with it and vice versa. CntnboolTEXTstopStopStopStopMsgeTEXTOnce finished with moving position of the bar and text, you can flatten the image using an action later in this set. This embeds the scale bar in your image so that the scale bar will change dimensions along with the image when re-scaled. CntnboolTEXTstopStopStopStopMsgeTEXTxDon't save over the original (this action has automatically duplicated your image to help prevent you from doing so). Black Scale BarTEXTstopStopStopStopMsgeTEXTcThis scale bar is NOT calibrated to your image! It is simply a bar with a length you can specify.CntnboolTEXTstopStopStopStopMsgeTEXTYou will need to know the pixel to micron (or other unit) calibration, something you can obtain by imaging a calibration slide with known values and then determining pixel to micron (or other unit) relationship.CntnboolTEXTstopStopStopStopMsgeTEXTYou will first be prompted to make a New file. Set the length of the New file in pixels to match your calibration. E.g., if 50 pixels = 100microns, enter 50 pixels. Do not change the width. that can be changed later. CntnboolTEXTmakeMakeMakeMk Nw ObjcDcmnNm TEXT white barMd typeGrysWdthUntF#Rlt@3333333HghtUntF#Rlt? =p RsltUntF#Rsl@rFl enumFl Wht TEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTfillFillFillFl UsngenumFlCnBlckOpctUntF#Prc@YMd enumBlnMNrmlTEXT copyEventCopyTEXTselectSelectSelectslctnullobj releDcmnTEXTpastePastePastepastAntAenumAnntAnnoTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTresetResetResetRsetnullobj propClr ClrsTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTstopStopStopStopMsgeTEXTThe next step allows you to transform or alter the width of the scale bar. Click on center box and drag to make smaller or larger. You will probably have to zoom in to make the bar larger.CntnboolTEXTstopStopStopStopMsgeTEXTWhen finished adjusting width, click on check icon at top, right of menu; or click on the marquee tool and then click "apply."CntnboolTEXT transform Transform TransformTrnfnullobj EnmrLyr OrdnTrgtFTcsenumQCStQcsaOfstObjcOffsetOfstHrznUntF#PxlVrtcUntF#Pxl?'΂HghtUntF#Prc@TH qTEXTstopStopStopStopMsgeTEXTNext, type in text for the unit of measurement. Highlight the portion of text you wish to change and type in new text. Move the position of the text by repositioning cursor away from text until cursor changes to"move" icon, then click and drag.CntnboolTEXTmakeMakeMakeMk nullobj ClssTxLrUsngObjcTxLr Txt TEXT25mmwarpObjcwarp warpStyleenum warpStylewarpNone warpValuedoubwarpPerspectivedoubwarpPerspectiveOtherdoub warpRotateenumOrntHrznTxtCObjcPnt HrznUntF#Prc@E GVrtcUntF#Prc@H5 yC textGriddingenum textGriddingNoneOrntenumOrntHrznAntAenumAnntAnCr textShapeVlLsObjc textShapeTEXTenumTEXTPnt TxttVlLsObjcTxttFromlongT longTxtSObjcTxtS$fontPostScriptNameTEXT Arial-BoldMTFntNTEXTArialFntSTEXTBoldScrplongFntTlongSz UntFC@4 syntheticBoldboolsyntheticItalicbool autoLeadingboolHrzSlongdVrtSlongdTrcklongAtKrboolBslnUntFCfontCapsenumfontCapsNrmlbaselineenumbaselineNrmlUndlbool strikethroughboolligatureboololdStyleboolproportionalNumbersboolRtt boolbaselineDirectionenumbaselineDirection withStreammojiZumedoub gridAlignmentenum gridAlignmentromanhyphenationLanguageenumhyphenationLanguageenglishLanguage wariChuCountlong wariChuScaledoub? wariChuWidowlong wariChuOrphanlongnoBreakboolClr ObjcRGBCRd doub@oGrn doub@oBl doub@oFl boolStrkbool fillFirstboolverticalUnderlinePositionenumverticalUnderlinePositionverticalUnderlineRightparagraphStyleRangeVlLsObjcparagraphStyleRangeFromlongT longparagraphStyleObjcparagraphStyleAlgnenumAlg LeftfirstLineIndentUntFC startIndentUntFC endIndentUntFC spaceBeforeUntFC spaceAfterUntFC hyphenateboolhyphenateWordSizelonghyphenatePreLengthlonghyphenatePostLengthlonghyphenateLimitlonghyphenationZonedoub@BhyphenateCapitalizedbooljustificationWordMinimumdoub?陙justificationWordDesireddoub?justificationWordMaximumdoub?G justificationLetterMinimumdoubjustificationLetterDesireddoubjustificationLetterMaximumdoubjustificationGlyphMinimumdoub?justificationGlyphDesireddoub?justificationGlyphMaximumdoub? hangingRomanbool burasagariboolpreferredKinsokuOrderenumpreferredKinsokuOrderpushIn mojiKumiNameTEXTNonetextEveryLineComposerboolautoLeadingPercentagedoub?33@ leadingTypeenum leadingType leadingBelowTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 1TEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTstopStopStopStopMsgeTEXTMove the position of the scale bar and text above it. These layers are linked, so if you move the scale bar the text will move with it and vice versa. CntnboolTEXTstopStopStopStopMsgeTEXTOnce finished with moving position of the bar and text, you can flatten the image using an action later in this set. This embeds the scale bar in your image so that the scale bar will change dimensions along with the image when re-scaled. CntnboolTEXTstopStopStopStopMsgeTEXTxDon't save over the original (this action has automatically duplicated your image to help prevent you from doing so).  White Bar & Text: Outline BlackTEXTstopStopStopStopMsgeTEXTXThis action will render the text layer so that it can't be edited. Just so you know...CntnboolTEXTselectSelectSelectslctnullobj propLyr BckgTEXThideHideHideHd nullVlLsobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXT mergeVisible Merge VisibleTEXTsetSetSetsetdnullobj propPrprLefxEnmrLyr OrdnTrgtT ObjcLefxFrFXObjcFrFXenabboolStylenumFStlOutFPntTenumFrFlSClrMd enumBlnMNrmlOpctUntF#Prc@YSz UntF#Pxl@Clr ObjcRGBCRd doubGrn doubBl doubTEXTshowShowShowShw nullVlLsobj propLyr Bckg Black Bar & Text: Outline WhiteTEXTstopStopStopStopMsgeTEXTXThis action will render the text layer so that it can't be edited. Just so you know...CntnboolTEXTselectSelectSelectslctnullobj propLyr BckgTEXThideHideHideHd nullVlLsobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXT mergeVisible Merge VisibleTEXTsetSetSetsetdnullobj propPrprLefxEnmrLyr OrdnTrgtT ObjcLefxFrFXObjcFrFXenabboolStylenumFStlOutFPntTenumFrFlSClrMd enumBlnMNrmlOpctUntF#Prc@YSz UntF#Pxl@Clr ObjcRGBCRd doub@oGrn doub@oBl doub@oTEXTshowShowShowShw nullVlLsobj propLyr BckgFlatten all layersTEXT flattenImage Flatten ImageSINGLE IMAGE LAYOUTImage Size + infoTEXTstopStopStopStopMsgeTEXTzThe image size dialogue box is poorly understood. Remember that the resolution is changed by checking "Resample Image." CntnboolTEXTstopStopStopStopMsgeTEXTThe image SHOULD be resampled when it is meant for publication in a journal or on the web. In that instance, set the resolution and the image dimensions and CHECK resample.CntnboolTEXTstopStopStopStopMsgeTEXTThe image should NOT be resampled if its intent is for printing or for PowerPoint: in that instance, only the dimensions need be changed. Resample box is NOT checked.CntnboolTEXT imageSize Image Size Image SizeImgSWdthUntF#Rlt@vRsltUntF#Rsl@rCnsPboolIntrenumIntpBcbc Image SizeTEXT imageSize Image Size Image SizeImgSWdthUntF#Rlt@vRsltUntF#Rsl@rCnsPboolIntrenumIntpBcbcMULTI-IMAGE AUTO-LAYOUTTEXTstopStopStopStopMsgeTEXTAll image files should be closed when using multi-image layouts. You will be prompted for which images to open. It is assumed that all images were already optimized.CntnboolTEXTstopStopStopStopMsgeTEXTdIf you encounter problems when running these actions, image files could be 16-bit or indexed color.TEXTstopStopStopStopMsgeTEXTIf you want 3 images across, then you can use a 6-panel portrait auto layout and repeat the same 3 images for the bottom row. Crop out the bottom 3.USA LETTER SIZETEXTstopStopStopStopMsgeTEXT\These auto-layouts will make the set of images as large as possible on an 8.5X11 inch page.CntnboolTEXTstopStopStopStopMsgeTEXTWhen the action is complete for each layout, these can be made to any dimension and any resolution by using Image Size (Image > Image Size). CntnboolTEXTstopStopStopStopMsgeTEXTtYou will need to have downloaded templates from the quickphotoshop.com download page in order to use these actions.2 panel topbottom (portrait)UTEXTstopStopStopStopMsgeTEXTOpen template first, then image files of interest. If the action stops midway with a prompt, an image is likely the wrong mode (indexed color or 16-bit). Change the mode first (Adjust > Mode).CntnboolTEXTresetResetResetRsetnullobj propClr ClrsTEXTopenOpenTEXTselectSelectSelectslctnullobj propLyr BckgTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTopenOpenTEXT flattenImage Flatten ImageTEXT$3caa3434-cb67-11d1-bc43-0060b0a13dc4 Fit ImagenullWdthUntF#Pxl@HghtUntF#Pxl@LTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdTpTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdLfTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTopenOpenTEXT flattenImage Flatten ImageTEXT$3caa3434-cb67-11d1-bc43-0060b0a13dc4 Fit ImagenullWdthUntF#Pxl@HghtUntF#Pxl@LTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTinverseInverseTEXT subtractFrom Subtract FromSubtract FromSbtFnullobj propChnlfselT ObjcRctnTop UntF#PxlLeftUntF#PxlBtomUntF#Pxl@RghtUntF#Pxl@TTEXT subtractFrom Subtract FromSubtract FromSbtFnullobj propChnlfselT ObjcRctnTop UntF#PxlLeftUntF#Pxl@{pBtomUntF#Pxl@RghtUntF#Pxl@TEXTmakeMakeMakeMk nullobj ClssLyr belowboolTEXTfillFillFillFl UsngenumFlCnBlckOpctUntF#Prc@YMd enumBlnMNrmlTEXTcutCutCutcut nullobj propChnlfselT ObjcOffsetOfstHrznUntF#PxlVrtcUntF#Pxl@DTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 2TEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr BTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdTpTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTdeleteDeleteDeleteDlt nullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 2TEXTselectSelectSelectslctnullobj nameLyr BTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdLfTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXT colorRange Color Range Color RangeClrRClrsenumClrsHghlTEXTdeleteDeleteTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTselectSelectSelectslctnullobj nameLyr BTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXT colorRange Color Range Color RangeClrRClrsenumClrsHghlTEXTdeleteDeleteTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXThideHideHideHd nullVlLsobj nameLyr Layer 2TEXThideHideHideHd nullVlLsobj nameLyr Layer 1TEXThideHideHideHd nullVlLsobj propLyr BckgTEXT mergeVisible Merge VisibleTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Nm TEXTtextTEXTshowShowShowShw nullVlLsobj nameLyr Layer 2TEXTshowShowShowShw nullVlLsobj nameLyr Layer 1TEXTshowShowShowShw nullVlLsobj propLyr BckgTEXThideHideHideHd nullVlLsobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr textTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTstopStopStopStopMsgeTEXT}You can now adjust the selection around your images. Click on small corner box and drag to adust (you may need to zoom in).CntnboolTEXTstopStopStopStopMsgeTEXTtDouble click inside the outline or click on the check mark at the upper right of the menu bar to complete the crop.CntnboolTEXTstopStopStopStopMsgeTEXTNote: if the lettering is not visible and you have a black background, it's because the lettering is also black. Change to white with action included in this set later on.CntnboolTEXT transform Transform TransformTrnfnullobj propChnlfselTEXTcropCropCropCropTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsaveSaveSavesaveCpy bool2 panel portrait RGB ColorVTEXTstopStopStopStopMsgeTEXTOpen template first, then image files of interest. If the action stops midway with a prompt, an image is likely the wrong mode (indexed color or 16-bit). Change the mode first (Adjust > Mode).CntnboolTEXTresetResetResetRsetnullobj propClr ClrsTEXTopenOpenTEXTselectSelectSelectslctnullobj propLyr BckgTEXT convertMode Convert Mode Convert ModeCnvMT typeRGBMTEXT duplicate Duplicate DuplicateDplcnullobj EnmrDcmnOrdnFrstTEXTselectSelectSelectslctnullobj releDcmnTEXTcloseCloseCloseCls SvngenumYsN N TEXTopenOpenTEXT flattenImage Flatten ImageTEXT$3caa3434-cb67-11d1-bc43-0060b0a13dc4 Fit ImagenullWdthUntF#Pxl@HghtUntF#Pxl@LTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdTpTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdLfTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLockingprotectPositionboolTEXTopenOpenTEXT flattenImage Flatten ImageTEXT$3caa3434-cb67-11d1-bc43-0060b0a13dc4 Fit ImagenullWdthUntF#Pxl@HghtUntF#Pxl@LTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXT copyEventCopyTEXTcloseCloseCloseCls SvngenumYsN N TEXTpastePastePastepastAntAenumAnntAnnoTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTinverseInverseTEXT subtractFrom Subtract FromSubtract FromSbtFnullobj propChnlfselT ObjcRctnTop UntF#PxlLeftUntF#PxlBtomUntF#Pxl@RghtUntF#Pxl@TTEXT subtractFrom Subtract FromSubtract FromSbtFnullobj propChnlfselT ObjcRctnTop UntF#PxlLeftUntF#Pxl@{pBtomUntF#Pxl@RghtUntF#Pxl@TEXTmakeMakeMakeMk nullobj ClssLyr belowboolTEXTfillFillFillFl UsngenumFlCnBlckOpctUntF#Prc@YMd enumBlnMNrmlTEXTcutCutCutcut nullobj propChnlfselT ObjcOffsetOfstHrznUntF#PxlVrtcUntF#Pxl@DTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr Layer 2TEXTlinkLinkLinkLnk nullobj EnmrLyr OrdnTrgtT VlLsobj nameLyr BTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdTpTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTdeleteDeleteDeleteDlt nullobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 2TEXTselectSelectSelectslctnullobj nameLyr BTEXTalignAlignAlignAlgnnullobj EnmrLyr OrdnLnkdUsngenumADStAdLfTEXTselectSelectSelectslctnullobj nameLyr ATEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXT colorRange Color Range Color RangeClrRClrsenumClrsHghlTEXTdeleteDeleteTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTselectSelectSelectslctnullobj nameLyr BTEXTsetSetSetsetdnullobj propChnlfselT obj EnmrChnlChnlTrspTEXT colorRange Color Range Color RangeClrRClrsenumClrsHghlTEXTdeleteDeleteTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXThideHideHideHd nullVlLsobj nameLyr Layer 2TEXThideHideHideHd nullVlLsobj nameLyr Layer 1TEXThideHideHideHd nullVlLsobj propLyr BckgTEXT mergeVisible Merge VisibleTEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr Nm TEXTtextTEXTshowShowShowShw nullVlLsobj nameLyr Layer 2TEXTshowShowShowShw nullVlLsobj nameLyr Layer 1TEXTshowShowShowShw nullVlLsobj propLyr BckgTEXThideHideHideHd nullVlLsobj EnmrLyr OrdnTrgtTEXTselectSelectSelectslctnullobj nameLyr Layer 1TEXTsetSetSetsetdnullobj EnmrLyr OrdnTrgtT ObjcLyr  layerLockingObjc layerLocking protectNoneboolTEXT mergeVisible Merge VisibleTEXTshowShowShowShw nullVlLsobj nameLyr textTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnAl TEXTstopStopStopStopMsgeTEXT}You can now adjust the selection around your images. Click on small corner box and drag to adust (you may need to zoom in).CntnboolTEXTstopStopStopStopMsgeTEXTtDouble click inside the outline or click on the check mark at the upper right of the menu bar to complete the crop.CntnboolTEXTstopStopStopStopMsgeTEXTNote: if the lettering is not visible and you have a black background, it's because the lettering is also black. Change to white with action included in this set later on.CntnboolTEXT transform Transform TransformTrnfnullobj propChnlfselTEXTcropCropCropCropTEXTsetSetSetsetdnullobj propChnlfselT enumOrdnNoneTEXTsaveSaveSavesaveCpy bool2 panel sidebyside (landscape)KTEXTstopStopStopStopMsgeTEXTOp