Difference between revisions of "Microphotography"

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-The article deals with '''microphotography''', i.e. 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'''.
-=Camera mode (settings)=
+=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.
@@ Line 8: / Line 29: @@
 *Normal.
-=Focusing=
+==Focusing==
 Setting live view:
 *Rench (II).
@@ Line 17: / Line 38: @@
 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=
@@ Line 41: / Line 73: @@
 #Set to "background color".
-==Edge effects/vignetting==
+==Vignetting==
-The lens distortion/lighting effects at edge of image are known as: "barrel distortion" and "vignetting".
+Defintion:
+*The edge of images are darker than the centre.
-Notes:
+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 (GIMP)===
+===De-vignetting===
+====Student method====
 #Take picture as one wants.
 #Remove slide and shoot the background.
@@ Line 58: / Line 100: @@
 #Invert background.
 #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==
@@ Line 65: / Line 122: @@
 *[http://hugin.sourceforge.net/ Hugin (sourceforge.net)].
-Image:
+===Images===
-*[http://commons.wikimedia.org/wiki/File:Londonpanorama.jpg Example of stitching (WC)].
+<gallery>
+Image:Londonpanorama.jpg | Example of stitching. (WC)
+Image:Enterobius - very low mag.jpg | Another example of stitching. (WC)
+</gallery>
 ==Focus stacking==
@@ Line 76: / Line 136: @@
 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=
@@ Line 81: / Line 157: @@
 *[[Digital ISO]].
 *[[Micrographs]].
+*[[Image annotation]].
+*[[Micrograph wishlist]].
 =References=
-{{reflist|2}}
+{{reflist|1}}
 =External links=

Difference between revisions of "Microphotography"

Microphotography (view source)

Revision as of 22:00, 2 January 2016

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