What’s Cooking In Microscopy:

A Microscopic Look at the Contents of A Refrigerator

Darkfield 40x Shark Muscle Cells

DAPI Stained Fluorescent Nuclei Within Shark Muscles

Adobe Photoshop: Adjust Contrast - Brightness

Combined ImageDarkfield and Fluorescent Microscopy Done in Layers with Adobe Photoshop

Besides their multinucleated status, shark muscle cells have another aspect, clearly visible the polarized and DIC image below. Both 60x images are of the exact same specimen and clearly show the striations present in shark muscle tissue. Obviously, the two techniques produced dramatically different results. DIC shows slight variations in the muscle surface, while the polarized image shows only the most dramatic aspects of the striations in sharp relief.

60x Polarizer: same image!60x DIC

Blue Nuclei - DAPI

Red Microtubules - Phallicidin

Green N-acetyl glucosamine residues in the cells’ surface - WGA

Incidentally, the Shark Tissue was triple-stained with fluorescent dyes (10x)

Phase 40 x

Recrystalized Salt, 40x:

Phony Hoffmann Effect – Light Microscope

Focus Through Layers, 100 x Brightfield, oil immersion

Red Leaf Lettuce

Adobe Photoshop Hue / Saturation Adjust

Phase

Bottom 2 Images: Angled Stage Setting

3 Video Camera Images of Re-crystallized Sugar, 40x

Phase Image Converted to 8 bit grayscale in image J

But an Inverted Look-up Table produces dim results!

In Photoshop, Contrast and Brightness Adjusted

Look-Up Table Applied To Crystallized Sugar

This Lookup Table Replaces 176 – 255 Red Values with 0.

It Creates a Turquoise Effect where Highlights Occurred in the Image for a colored Look at Sugar’s Crystalline Patterns

Onion Cell Nuclei Stained With Iodine - based Fluorescent Stain

Random Confocal Slices – Indistinct, Noisy Images…

100 x Magnification

To the left: An Averaged Z-projection in Image J compiling all of the confocal slices. It produced a much cleaner image.

And with a little doctoring in Photoshop using

the diffuse glow filter and

the RGB channel mixer, the nuclei and cell membranes become much more distinct

Brightfield 40 x Image of Same Onion Skin, a little dull …lots of visual goo in the sample.

To the right:

A simple convolution applied adds a little interest:

0 1 2

-1 1 1

-2 -1 0

But Holy Cow!

The convolved onion skin jpeg doctored in Photoshop using Image>Adjust>Curves to give the cell membrane a psychedelic look , crazy man.

1. Image J: Image>Adjust>Brightness / Contrast

2. Image J: Process>Noise>Despeckle

3. PhotoShop: Filter>Smart Blur

4. PhotoShop: Filter>Noise>Despeckle

5. PhotoShop: Touch-up with Black and grey “paint” w/paintbrush6. PhotoShop: Filter>Noise>Add Noise

Fixing Gretchen…

The End