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The article deals with '''microphotography''' - creating microphotos.
[[Image:Teratoma 2 low mag.jpg|thumb|350px|[[Micrograph]] of a [[teratoma]]. [[H&E stain]].]]
The article deals with '''microphotography''', i.e. creating '''microphotos'''.
 
=Taking the picture=
==The keys to good pictures==
*Spend time on set-up.
**Clean the slide.
**Look for the pristine areas without artifacts, e.g. folds.
*Composition.
**Put normal beside pathologic - so one has a reference point.
***Transititions are usually interesting.
**Where to put the centre of interest:
***Truly beautiful: use the ''rule of thirds'' - see ''[[Microphotography#Composition|composition section]]''.
***Functional: centre of interest in the centre.
*Focus.
**Sharpness of nuclear membrane and detail in the cytoplasm.
*White balance.
*[[De-vignetting]].
*Pictures at different magnification.
**Sets are usually better than one.
**Many entities have high and low power features.
***It is often impossible to capture them with one picture.
 
==Camera settings==
Exposure compensation:
*None.
 
White balance:
*Normal.
 
==Focusing==
Setting live view:
*Rench (II).
**Live view function settings.
***Live View shoot.
*Press "Set" button to open the shutter.
 
Use the zoom button (on the camera): press twice to digitally magnify 10x.
*Focus microscope as usual (@ 10x digital magnification).
 
Note:
*Post-processing will not fix a blurry image.  The way to get sharp images is take sharp pictures!
 
==Composition==
[[Image:Rivertree thirds md.gif|thumb|150px|A picture that uses the ''rule of thirds''. (WC)]]
Functional pictures that tell a story and are easy to understand:
*The centre of interest at the centre.
 
An artsy look can be achieved by making use of the [[Wikipedia:Rule_of_thirds|rule of thirds]].
*Rule of thirds: centre of interest is at one of the four intersects of the imaginary lines that divide the image into thirds.
 
=Post-processing=
GIMP scripts:
*White-balance.
*Shadows & highlight.


==White balance==
==White balance==
Done with a GIMP plugin.<ref>URL: [http://registry.gimp.org/node/72 http://registry.gimp.org/node/72]. Accessed on: 26 July 2010.</ref>
*May be done with:
**White balance plugin.<ref>URL: [http://registry.gimp.org/node/72 http://registry.gimp.org/node/72]. Accessed on: 26 July 2010.</ref>
**''Curves'' function (in GIMP).
 
===Curves function (in GIMP)===
*Adjust magnitude, then blue, then green -- red should be last.
**Low magnification images tend toward "too pink" with the white-balance script.


===Actions===
===White balance plugin (script)===
Procedure:
#Select
#Select
#*Filters.
#*Filters.
Line 12: Line 73:
#Set to "background color".
#Set to "background color".


==De-vignetting==
==Vignetting==
Defintion:
*The edge of images are darker than the centre.
 
Microscope configuration:
*[[Köhler illumination]].
*Ensure diaphragm is open.
 
Some random notes:
*Software for microscope cameras usually have de-vignetting tools.  The tools go by different names and usually entail taking a background image.  Ideally, one should shoot the background image at the same magnification as the (primary) image.
*Vignetting is usually worse at lower magnification.
 
Random links:
*[http://www.gimp.org/release-notes/gimp-2.4.html http://www.gimp.org/release-notes/gimp-2.4.html].
*[http://www.gimp.org/release-notes/gimp-2.4-videos.html http://www.gimp.org/release-notes/gimp-2.4-videos.html].
 
===De-vignetting===
====Student method====
#Take picture as one wants.
#Take picture as one wants.
#Remove slide and shoot the background.
#Remove slide and shoot the background.


In GIMP:
In GIMP:
#Load background.
#Load image.
#Load image as a layer.
#Load background as a layer ("Open as layer... ").
#Make sure the background image is the top layer.
#Make background into an "overlay" in Layers dialog box.
#Make background into an "overlay".
#Invert background.
#Invert background.
#Save as jpg.
#Save as jpg (merge with background).
 
====Adapted from codeforhire.com====
Procedure:<ref>URL: [http://codeforhire.com/2013/06/29/simple-image-devignetting/ http://codeforhire.com/2013/06/29/simple-image-devignetting/]. Accessed on: October 16, 2014.</ref>
#Load image in GIMP.
#Duplicate layer (right click on layer and select ''Duplicate Layer'').
#Apply Gaussian blur to top layer.
#*Filters -> Blur -> Gaussian Blur
#**Select a ''blur radius'' 1/5 of the smallest dimension of the image.
#***If the image is 2768x2110, the blur radius should be 2110/5=422.
#Invert top layer (Colors - Invert).
#Change top layer to ''Overlay'' mode.
#Normal top (masking) layer (Colors - Auto - Normalize).
#Adjust brightness of (top) layer (Colors - Brightness-Contrast).
#Merge layers.
#*Right click on top layer and select ''Merge Down''.
 
==Stitching==
*A technique to increase the field of view.
 
Software:
*[http://hugin.sourceforge.net/ Hugin (sourceforge.net)].
 
===Images===
<gallery>
Image:Londonpanorama.jpg | Example of stitching. (WC)
Image:Enterobius - very low mag.jpg | Another example of stitching. (WC)
</gallery>
 
==Focus stacking==
*A technique to increase the [[depth of field]].
 
Software:
*[http://hugin.sourceforge.net/ Hugin (sourceforge.net)].
 
Image:
*[http://commons.wikimedia.org/wiki/File:Focus_stacking_Tachinid_fly.jpg Example of focus stacking (WC)].
 
==Dirt and defects==
*It is best to clean the slide.
**''Photons are free... time is not.''
*Dirt and defects can be removed with the ''clone'' tool in GIMP.
 
==Sharpening==
*Images should be sharpened to enhance edges. 
**This is particularly important if the image is projected on a large screen and/or enlarged.
**Over-sharpening makes images look like caricatures.
*Sharpening should be the last step in post-processing.
 
==Animation==
[[Image:Squamous carcinoma - lung FNA -- high and very high mag - animation.gif|thumb|right|An animated GIF showing [[squamous carcinoma of the lung]]. (WC)]]
*GIMP has a [https://www.gimp.org/tutorials/Simple_Animations/ nice tutorial (gimp.org)] on how it can be done. 
*Animations are particularly useful for [[cytopathology]].
 
=See also=
*[[Microscope]].
*[[Digital ISO]].
*[[Micrographs]].
*[[Image annotation]].
*[[Micrograph wishlist]].
 
=References=
{{reflist|1}}
 
=External links=
*[http://www.martinmicroscope.com/ Martin Microscope company (martinmicroscope.com)].


==References==
[[Category:Basics]]
{{reflist|2}}
[[Category:Microphotography]]
Michael
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