Live Blood Analysis: Leukocytosis

We were testing out some blood specimens yesterday we used a methylene blue I was testing some mountants for peripheral blood smears.
Stain used: methylene blue, polychromed by boiling with Na2CO3 for about 30 minutes. The smear was heat-fixed, brought to alcohol and then xylene, and finally covered with mounting medium and a cover slip. Thick smear leukocytes

 The CMOS eyepiece camera is a great way to get micro-critters onto a video screen, but it doesn’t show quite as much detail as the human eye can see looking through the eyepiece. However, you can see from the photo that there appears to be a mix of immune-response cells.

 On another note, you will notice there are no erythrocytes visible. The polychromed methylene blue is normally treated with eosin to give a Romanowsky stain; this reveals both red and white cells. The white cells’ cytoplasm collapsed or degraded severely in both test specimens after about a week, except in those areas where it was apparently stabilized  in the thicker regions of the smear.

The History of the Microscope

During that historic period known as the Renaissance came the invention of the light microscope. A revolutionizing instrument that enabled the human eye to observe microscopic objects. It enabled people to understand the fascinating details of the known world.

In 1590, two Dutch spectacle makers discovered that nearby objects appeared greatly enlarged. That was the forerunner of the compound microscope and of the telescope. In 1609, Galileo, father of modern physics and astronomy, heard of these early experiments, worked out the principles of lenses, and made a much better instrument with a focusing device.

The father of microscopy, Anton van Leeuwenhoek, started as an apprentice in a dry goods store where magnifying glasses were used to count the threads in cloth. He taught himself new methods for grinding and polishing tiny lenses of great curvature which gave magnifications up to 270 diameters, the finest known at that time. He was the first to see and describe bacteria, yeast plants, the teeming life in a drop of water, and the circulation of blood corpuscles in capillaries. He studied both living and non living, and reported his findings in over a hundred letters to the Royal Society of England and the French Academy.

Anton van Leeuwenhoek Microscope

(Via birthstory.com)

Robert Hooke is considered the English father of microscopy because he confirmed Anton van Leeuwenhoek's discoveries of the existence of tiny living organisms in a drop of water. Hooke made a copy of Leeuwenhoek's light microscope and then improved upon his design.

Robert Hooke Microscope 

(Via http://micro.magnet.fsu.edu/primer/museum/hooke.html)

A Microscope Master Review

Review: 

OMAX 40X-2000X Digital Binocular Biological Compound Microscope with Built-in 3.0MP USB Camera and Double Layer Mechanical Stage

"The construction and all round capabilities of this microscope surpass what you would normally expect for this price. 

 This Omax microscope offers affordability, convenience and durability. Images obtained are bright and clear, controls are firm and the stage and slide holder are easy to work with. 

 Equipped with a 3.0M pixel built-in USB camera, you will enjoy not only photo capability but video as well which is especially beneficial where participation in a group setting is necessary. 

 Pictures can be brightened, cropped and saved easily. 

 It is compatible with MS Windows 2000/XP/Vista as well as Windows 7 -32bit and 64bit. 

 Worth noting is Omax's excellent customer service and informative technical support."

Shop Model: M827S-C30

Cell Staining Simulation Tool

We recently discovered an amazing new tool from www.invitrogen.com.

Stain your own cell by using cell staining simulation to develop amazing results with their florescent dyes. 

Try it here: http://bit.ly/blBLo5

Before Stains

After Applied Stains

Living Cell Microscopy

Live cell microscopy is now a very common form of research in many fields in life science and other physical sciences. Live cell microscopy is crucial when performing experiments where cell visibility is at the center of measurement to ensure that biological processes are not altered in anyway.

 

Most cells and tissues are never exposed to sunlight so proper microscopy procedures that minimize light exposure. To ensure minimal light exposure, microscopes should be optimized to collect as much of the light source as possible.

 



 

 

It is crucial to ensure suitable environmental conditions on the microscope stage to maintain living cells. With an efficient optical microscope and detector, the light exposure can be minimized during live cell imaging, thus minimizing phototoxicity.

 

Perfect for Live Blood Analysis:

5.0 MP Digital Darkfield Siedentopf PLAN Microscope

(40X-1600X) $1,199.99

Shop here

Uses for Darkfield Microscopy

Darkfield microscopy allows for beautiful, detailed images to be revealed with one opaque disk used to block the light into just a few scattered beams. The background is dark and the sample reflects the light of the beams only. This results in a light colored specimen against a dark background. Darkfield microscopy is perfect for viewing clear or translucent details.

When to Use a Darkfield Microscope:

• Single- celled organisms
• Live Blood Analysis
• Transparent Specimens
• Live Bacteria
• Soil Samples
• Pond water Bacteria
• Seawater Samples
• Pollen Samples

Darkfield microscopy make most invisible specimens appear visible under the microscope, especially living organisms. 

Shop Darkfield Microscopes: http://bit.ly/11Rl53o

How to Set Up a Phase Contrast Microscope

Phase contrast is a microscopy technique that is helpful in viewing many biological specimens such as bacteria or blood cells. When setting up your phase contrast microscope, you must have phase contrast objective lenses and a phase contrast condenser.

The condenser shown above has five settings on it: 10x, 20x, 40x, 100x and bright field. The two screws that stick out from the condenser are centering screws and will be used when you set up the phase contrast microscope for the first time.  Set the condenser on the bright field setting and focus on a specimen. Adjust the height of the condenser for optimum image quality. Move the condenser turret to the phase setting for whichever lens you are currently using and remove the specimen.

Remove one of the eyepiece lenses and insert the centering telescope in its place. If there is a set screw on the side of your centering telescope this should be used to focus the centering telescope. When looking through the centering telescope you will see two rings. By turning the centering adjustment screws on the condenser, you can align the rings so they are concentric.

Remove the centering telescope and replace the eyepiece lens. Put your specimen back on the stage and you are ready for phase contrast observation! You will want to go through this process when you change objectives.

Shop Phase Contrast Kits: http://bit.ly/11Rn0yh

Ultra Violet Lights

Ultraviolet light also referred to as UV light can be used as a germicidal agent to sterilize surfaces where sensitive microbiology techniques are being performed. UV light is non-ionizing short wavelength radiation that falls between 4-400 nano-meters in the visible spectrum.

In general, the shorter the wavelength the more damaging it is to cells which makes UV light more damaging than visible light or infrared light. Most bacteria are killed by UV light. UV radiation at 260 nm is most damaging because at this wavelength DNA maximally absorbs UV light.

When DNA absorbs UV light pyrimidine dimers form.  These dimers distort the overall structure of the DNA strand and prevent DNA polymerase from moving past the dimer. Genes downstream from the dimer will not be transcribed and essential proteins will not be formed.

The ability of the organism to function normally will be affected by the formation of one dimer. Ultraviolet light kills cells by damaging their DNA. The light initiates a reaction between two molecules of thymine, one of the bases that make up DNA.  Even so, it breaks down when the damage is extensive.