Fluorescence Microscopy Tips

The images you take are only as good as your equipment. So take time to take care of your equipment on a regular basis; clean objectives, cover slips, and ensure your microscope has the proper setup.

Most objectives are designed for a cover-slip that is .17mm thick. If you fail to use the proper cover-slip, you will lose image clarity and sharpness. Although cover-slip thickness can range anywhere from .08mm to .25mm in thickness, .17mm is the most common and optimal for most microscopes.

Slides are also another component that vary in quality. Research your brands and find out which is the best for your type of microscope.

For further tips we recommend referencing the following text:
"A Guide to Super-Resolution Fluorescence Microscopy" (2010) Schermelleh et al. J Cell Bio.
"Live Cell Microscopy- Tips and Tools" (2009) Frigault et al. J Cell Sci.

Objective Lens Comparison

Objective Lenses: 
Achromatic vs Semi-Plan vs Plan 

Besides some special microscope objective lenses, there are three kinds of objective lenses on the market.
Achromatic, semi-plan, and plan. 

Achromatic Lens:

Semi-Plan Lens:

Plan Lens: 

 

First of all, all these three objective lenses correct for color, although there's no "achromatic" in the name of "semi-plan" or "plan". The difference between these three lenses is the focusing area which can be seen from the eyepieces when using high power objective lenses such as 40x, 60x and 100x. 

The difference from low power lenses are not obvious, which means that can be ignored. At high power, an achromatic lens has about 65% of focused area acrossing the center. A semi-plan one has about 85% whereas the plan one has 95%. 

 Due to the correction from the different lenses, the price for those lenses are also different. Here are some links for the objective lenses: 

Achromatic: http://www.microscopenet.com/achromatic-objective-lenses-compound-microscopes-p-377.html 
Semi-Plan: http://www.microscopenet.com/semi-plan-achromatic-objective-compound-microscopes-p-317.html
Plan: http://www.microscopenet.com/plan-achromatic-objective-compound-microscopes-p-316.html

Objectives

The following is a list of descriptions of the various types of objectives available for your microscope.

 Achromat Objectives - these are the most basic high power microscope objectives. Typically the outer edge of the circular image seen through the microscope will be slightly out of focus due to the curvature of the lens.

Semi-Plan Achromat Objectives - these are one step up from basic microscope objectives in that the field of viewhas been corrected a bit and the outer edges will be in focus more than the standard achromat lens.

Plan Achromat Objectives - these microscope objectives have been corrected for lens curvature and the entire image plane is flat and in focus

Plan Apochromat Objectives - These objectives have been corrected for four colors chromatically and spherically. Best used for critical resolution and color photomicrography.

Plan Fluorite Objectives - these microscope objective lenses are corrected for four wavelengths, but not quite as extensively as plan apochromat objectives.

100 X Achromatic Compound Microscope Objective Lens $69.99
Shop here: http://bit.ly/130Jfs8http://bit.ly/130Jfs8

Shop Objectives: http://www.microscopenet.com/microscope-accessories-objectives-c-89_53.html

DIY: Slides

A great project for kids of any age is making a variety of microscope slides to view under your high power microscope.

When making temporary slides, the supplies needed are very basic:
1. blank slide
2. cover slip In order to make your slide, simply place the specimen on the slide, apply the cover slip so it lays flat, and view under the microscope.
3. When finished viewing, wash the slide in warm soapy water. The cover slip is quite thin so be careful that you don't break it during cleaning.

Creating permanent slides involves sealing your specimen within the slide so it lasts, and then labeling the slide so that years from now you remember what the sample was. You may also want to stain the slide, if so, this page offers great information on staining microscope slides.
1. Prepare the slide just as you would when making temporary slides.
2. When applying the cover slip, you will want to coat the very outer edges of the cover slip with a thin layer of clear nail polish or cement in order to permanently seal the cover slip to the slide.
3. Make sure your layer of adhesive is not too thick, as this will cause problems with not being able to focus properly on your specimen.

Fun slide making ideas!
1. Cheek cells from inside your mouth (use a Q-tip to scrape the inside of your mouth).
2. Pond water, or water from a muddy puddle.
3. Fine sand particles, sugar, salt, baking soda, a vitamin C tablet - any fine powdery substance. (If you have a polarizing filter - use it when viewing these items as well!)
4. Fabric fibers or strings. Cotton - a thin strand pulled off a cotton ball.
5. Soil from the back yard. A blade of grass or a thin section of a flower.

Adjusting Condensers

The condenser aperture diaphragm is used to control the contrast and resolution of an image. An improper setting of the condenser aperture diaphragm  can be the cause of much frustration both for teachers and students. Students may attempt to find the focus with the condenser aperture diaphragm all the way open.

Remember, an open condenser aperture diaphragm results in a low depth of field. Students may not see anything at all when working with high magnifications because the image is too dark. In this case the diaphragm is closed too much. The diaphragm should not be used to control the amount of light, but for some specimens or magnifications there may simply be no way around this especially if the lamp is not very powerful. The higher the magnification, the more need for a condenser.

Tips:

• Completely close the condenser aperture diaphragm when starting to use the microscope. 
• Rotate the low power objective into position and find the focus with the coarse focus knob. The larger depth of field and higher contrast makes it easier for the students to focus the specimen. 
• When switching to a higher magnification, the students should start to gradually open the condenser aperture diaphragm, to observe the differences in image quality. 
•  Adjust the light intensity with the dimmer to prevent glare. Students should be made aware that the condenser aperture diaphragm should be adjusted to the numerical aperture value which is printed on the objective.
• Opening the diaphragm further will not increase image quality, but may result in glare. If the sample is thick, strongly stained or pigmented then the diaphragm has to be opened to allow more light to pass through the specimen. As a consequence, the depth of field becomes smaller.
•  Use the fine focus adjustment knob to focus through the different layers of the specimen

Shop Darkfield Condensers here: http://bit.ly/16rya5k

Gem Microscopes

A gem microscope is similar to a biological or medical microscope in that it is binocular, and uses compound lenses. 

A binocular magnifying device has two eyepieces so that both eyes are used at once. This is ideal for getting a good three dimensional view. In a compound scope, there is a set of lenses close to the object being magnified and a set in the eyepieces.
With this set-up, magnification is compounded, meaning that, for example, if the objective lens is 5x and the ocular lens is 10x, the total magnification is 50X (5X x 10X).

Gem scopes differ from biological scopes in that the total maximum magnification is usually lower and there are more lighting options. 

Professional grade gem microscopes generally include:

• brightfield illumination
• darkfield illumination
• oblique fighting
• overhead lighting
• light diffusing system
• a system for immersing the object in liquid in a well for viewing
• polarizing lighting
• pivoting stone holder

Shop our selection of Jewel Gem Microscopes here.

Check out our Professional Gem Jewel Darkfield Stereo Microscope 12X-75X

On sale now for $1,244.99

Shop Microscope here

Gemology: Diamonds

Diamonds are one of our links between the world back then and the world now. We don't know if they truly are forever but we do know that they have been in existence a very long time.

Diamonds are made of carbon atoms that are formed into a tightly packed structure with extreme heat, pressure, and oxygen conditions. Most diamonds form in the earth's crust or mantle but some deeper down. Under the earth's crust they form as impact diamonds under the deep earth. due to mass pressure. The redox reaction of CO2 or Methane forms diamonds. The complex pattern of diamonds could take anywhere from 3 billion years to form in the earth's mantle and brought so the earth's surface through magma.

Shop Jewel Gem System Microscopes here

Darkfield Imagery Tips

Darkfield microscopy is a very simple method for viewing unstained specimens clearly because it is clearly visible. Good candidates for darkfield observation usually have a transparent background that way the light an illuminate the specimen and darken the surrounding area, making the image quality clearer and more intense. Good specimens to use include aquatic organisms, cells, and cultures.

To achieve good darkfield images make sure that the central light rays along the optical axis are blocked. Blocking these light rays allows for just oblique rays to pass through at sharp angles, striking the specimen and illuminating the image. A darkfield condenser will get you the results your looking for with your microscope. The condenser reflects a single surface that provide more even illumination.

We recommend the OMAX Compound Binocular Microscope with Darkfield Condenser and 5.0 MP Digital Camera for $549.99

Model No: M827-A191-C50

CLICK HERE TO SHOP

Compound Microscopes 101

A compound microscope can be used for medical research to a day at the beach. A compound microscope includes an eyepiece, stage clips, and objectives with different lenses, adjustable knobs, a power button, and a stage with light source. Other parts include the body, nose piece, arm, and stage stop.

It can be used in various science fields such as Microbiology and Geology. Forensic investigators and scientists identify crystal shapes to reference pharmaceuticals and other medical needs. Microscopes can also help them see human cells, minerals, and metals. 

The setup of the optical parts plays an important ole in working with a compound microscope. The condenser focuses light onto the specimen with either bright-field or darkfield condenser attachments. 

By understanding these basics, you can independently carry out an experiment for observing minute objects of your interest, and try to explore some of the incredible things with the help of this wonderful instrument. Nowadays, compound microscopes are used in various domains, within or outside science. Care should be taken while buying any of the microscopes, be it compound or digital microscope. For better understanding, you can always discuss with a science teacher or an expert.

For more info check out our website's shopping guide here.

Shop Compound Microscopes here.

What is Phase Contrast Microscopy?

Phase contrast is a microscopy method used by Fritz Zernike. Zernike made multiple discoveries including the direct path of light. Phase contrast microscopy makes images appear darker against a light background.  Phase contrast uses a condenser system and an objective lens. Phase is only useful on images that are colorless or transparent because it makes it difficult to see it's surroundings. Objects suitable for phase contrast microscopy are bacteria, protozoa, and cells. Zernike was awarded the Nobel Peace Prize in 1953 for his discoveries in Physics.

We recommend our 2000X Phase Contrast Compound LED Microscope with 9.0 MP Camera.
Shop here: http://bit.ly/14WLZrU

Product Features Include:

• High Quality Glass Elements
• High Definition Images
• Eight Levels of Magnification
• USB Camera
• Advanced Software
• Abbe Condenser

OMAX MicroscopeNet Facebook Promotion

We at MicroscopeNet are hosting our first ever Facebook promotion where you can enter for your chance to win a microscope just by "liking" our Facebook page!

Here's how to enter to win:
Step 1: Like Microscopenet on Facebook, then go to "Enter To Win" tab.
Step 2: Once you have completed an Enter Form, then you must submit your Entry Form at our Facebook page.
Step 3: Share this event with your friends, and you can get 1 bonus entry every time a friend enters!  

If you are the lucky winner you will have the choice of one of our two most popular models below. No return for cash value. You are entitled to our 5 year warranty on the microscope you win. Free UPS ground shipping for continental US and Canadian address. AK,HI, and where UPS Ground does not reach, and international winner will have to pay for the extra shipping fee.

You can choose from the:
1) Binocular Stereo Microscope with Dual LED Lights

• Plug and see, no special knowledge needed
• High quality optical glass elements
• Three magnification levels: 20x, 40x & 80x
• 45°inclined 360°rotatable binocular head
• Dual LED lights (upper and lower)
• LED light lifetime: 100,000 hours
• Long working distance
• Sharp stereo erect images over a wide field view
• Diopter adjustment on left ocular-tube
• Adjustable inter-pupillary distance
• 90°rotational objective
• Locked-in eyepieces
• Tool-free focus knob tension adjustment
• Metal stand and framework

2) Compound Binocular Biological Microscope with Replaceable LED Light

• 8 levels of magnification: 40X-80X-100X-200X-400X-800X-1000X-2000X
• Replaceable 3W LED transmitted light with adjustable intensity
• 45 degree inclined 360 degree swiveling binocular viewing head
• Large double layer mechanical stage with NA1.25 Abbe condenser & iris diaphragm
• Coaxial coarse & fine focus knobs with tension control

Click here to "like" our Facebook page

Illumination Types for Microscopes

• Tungsten – the least expensive method, and the most common on low-end scopes, tungsten illuminators use standard incandescent light bulbs. They are relatively bright, but they produce a yellowish light and considerable heat. In particular as the light is dimmed, it shifts further toward orange. This warm color balance can obscure the true colors of specimens. The heat produced by the incandescent bulb may kill live specimens and quickly dries out temporary wet mounts made with water. Lamp life is relatively short.

• Fluorescent – costs a bit more than tungsten, and was quite popular before the advent of LED illuminators. Fluorescent illuminators provide bright light that appears white to the human eye, but is actually made up of several different discrete colors that are mixed to appear white. Accordingly, color rendition can differ significantly from the true color rendition provided by daylight. Fluorescent bulbs emit much less heat than incandescent bulbs, and so are well suited to observing live specimens. Some fluorescent illuminators are battery-powered, but most use AC power. Lamp life is relatively long.

• LED – priced about the same as fluorescent illuminators, LED illuminators have become very popular, largely replacing fluorescent illuminators. LED illuminators have the same color-rendition problems as fluorescent illuminators, but are otherwise ideal for many purposes. LED illuminators draw very little power and emit essentially no heat. Their low power draw means they’re the best choice for a battery-powered microscope, and are ideally suited for portable microscopes that can be used in the field. Lamp life is essentially unlimited.

• Quartz-halogen – the most expensive type of illuminators  and the one preferred by most scientists. They provide a brilliant white light needed for work at high magnification that reveals the true colors of specimens. Unfortunately, quartz-halogen lamps also produce more heat than any other type of illuminators  Their high power draw means they are AC-only. Lamp life is relatively short.

Cleaning Your Microscope

Caring for your microscope is essential to keep all the components working smoothly and maintaining the cleanliness and appearance of your microscope will help ensure clear visibility when looking through your eyepiece. 

It is best to only clean the eyepiece when needed. Use the proper cleaning materials which can be purchased on our website under the accessories category or from another independent retailer. If your eyepiece needs cleaning you can dip your cloth into an alcohol solution to remove stubborn spots.

The eyepiece and objective lens can obtain build up easily from use. If your objective lens is also dirty you can clean it with lens paper dipped into a diluted ammonia solution. Place a few drops of ammonia into about 1/2 cup of water. 

To ensure you will not scratch your eyepiece(s) or lenses refrain from using anything other than a special lens cleaning cloth or tissue. Do not use your finger nails to try to remove any spots or debris.
Click here to browse our selection of cleaning tissue. 

Trinocular Zoom Stereo Microscopes

Trinocular Zoom Stereo Microscopes are amazingly useful for lab research and hobbyists alike. With the third tube in place, you can easily hook up with the usb camera to your TV or PC monitor.  No mather what you're inspecting for, coin, rocks, or gems our selection of trinocular stereo microscopes won't disappoint. Stereo Microscopes are practical for all levels of observations.  They are a delight for gemologists, jewelers, collectors, and even hobbyists. We recommend our Trinocular Zoom Stereo Microscope with 8W Fluorescent Ring Light and 9.0 MP Camera on sale now for $725.99! It's great for larger specimens because it comes with a table stand and can easily be connected to your laptop for still and live image recording.  Product Features Include: High quality optical glass elements 2 pairs of widefield eyepieces (10X and 20X) 0.5X auxiliary objective lens provides larger field of view and longer working distance 8W fluorescen ring light provides shadowless illumination 9.0 MP digital camera

How to Focus a Microscope

Focusing your microscope is the key to getting the clear image quality you expect out of your microscope. When first looking at a specimen, the item will almost always never be focused. Learning how to focus your microscope is the essential step to assure proper observations and results. 

Here a five easy steps for focusing your microscope:
1. Place prepared slide underneath the viewing tray of the microscope and secure with clips 

2. Look through your eyepiece and determine if you can see the specimen. The specimen may be fuzzy and/or unrecognizable.

3. Turn the coarse focus knob slowly. Continue turning the coarse focus knob until the specimen becomes recognizable.

4. Next slowly turn the fine focus knob until the image becomes sharper and clear. At this point small details about the the specimen should be crystal clear. 

5. Adjust the diaphragm of the microscope to ensure the entire specimen is in your peripheral view. 

Featured Product: Student Microscope 40X-2000X Lab Binocular Biological Compound Microscope

A compound microscope is ideal for students majoring in bio/medical fields as well as for homeschooling and hobbyists. Compound microscopes are ideal for slide observations, which is what most students are required to examine in the classroom. Our Binocular Biological Compound Microscope is an ideal starter microscope for students and is on promotion right now for a low price of $198.99. 

This microscope is great for students because it can be easily upgraded with a digital camera, a darkfield adjustment, and slides. This microscope is easily customizable and easy to use. Compound microscopes will continue to be a crucial part of research in scientific fields. The compound microscope is still a mainstay in many fields and also is a wonderful tool for students to begin with when learning how to use these instruments in a laboratory. Their ability to provide higher magnifications and excellent resolution of a specimen makes the compound microscope an important tool for scientists and students alike.

Product features include: 

• High quality professional optical glass elements
• 45°inclined 360°swiveling binocular head
• Easy to mount electronic eyepiece (digital microscope camera)
• 8 levels of magnification: 40X-80X-100X-200X-400X-800X-1000X-2000X
• 4 achromatic objectives 
• 2 pairs of eyepieces: WF10X and WF20X
• Sliding interpupillary distance adjustment
• Ocular diopter adjustable on both eyetubes
• Variable intensity illumination
• Coaxial coarse and fine focus adjustment
• Focusing knobs on both sides
• Stage upward moving lock protects objectives and slides
• Stain-resistant double layer mechanical stage
• NA1.25 Abbe Condenser with iris diaphragm & filters
• Rack and pinion adjustment condenser
• Product GS and CE approved

More on Darkfield Microscopy

Darkfield microscopy requires a special stand containing a reflective mirror and light shielding plate at a specific angle. The inverted hollow cone is projective of the desired image at an oblique angle. This process is the same for both stereo microscopes and standard compound microscopes. 

Good specimens for darkfield microscopy are those close in value to their background images. Small aquatic organisms and cells are ideal for such microscopic lighting technique. Specimens preferred for use with brightfield lighting are not recommended. Other ideal specimens for darkfield microscopy under a stereo microscope include bones, fibers, yeast, and hairs.

Illumination from brightfield lighting involves blocking out central light rays along the central axis. Blocking these rays allows for only oblique angled rays to strike the specimen resulting in a hollowed cone illumination of the desired specimen.

In darkfield lighting, contrast is greatly enhanced by the lighting technique at hand. Highly oblique light rays, diffracted by the specimen and yielding first, second, and higher diffracted orders at the rear focal plane of the objective, proceed onto the image plane where they interfere with one another to produce an image of the specimen.

We recommend our 1600X Darkfield Compound 9MP Digital Siedentopf PLAN Microscope
with 4 PLAN objectives and darkfield condenser. 

Featured Product: Trinocular Zoom Stereo Microscope

This week we are featuring one of our top selling microscopes..
Our 7X-45X Trinocular Zoom Stereo Microscope with Dual Lights includes a 5.0 Megapixel Camera and software compatible with Windows XP/Vista/7. The included software can be used to make measurements on your PC. The 2592 X 1944 pixels from the USB digital camera can capture both still and live images on your computer, making this microscope ideal for live specimen analysis and laboratory observations.

This microscope includes an adjustable barlow lens that can extend the working distance up to 6.25" long and the field of view to be as wide as 2.38" long. This microscope also includes dual halogen lights with adjustable intensities.  This microscope is ideal for biological research labs, engravers, watchmakers, gemologists, jewelers, collectors, and those in electronic related industries. 

This microscope is also easily customizable as well. You can add a darkfield stage for darkfield observations. A boom stand and gooseneck light for a larger work space. You can also add different eyepieces, lights, and cameras. The possibilities are endless with the Trinocular Zoom Stereo Microscope. 

Shop Microscope now: Click here

Microscopes for Homeschoolers

Microscopes are beneficial for students who are homeschooled because it enables them to have the same type of exposure to science and experiments as those enrolled in public education institutions. 

 

The first thing you need to decide is what kind of observations your child/student will be expected to view and what kind of experiments he/she will partake in during the school year. Next you will need to decide if it is best to purchase a compound or stereo microscope. 

A compound microscope is only useful got looking at slides, which may get rather boring for students, slides must be purchased or made at home, which can be tedious and time consuming. A stereo microscope will enable your student to view live specimens, 3-D objects, and some slide use. 

For normal observations, you probably will not need a microscope with a magnification higher than 400X. 400X magnification is generally the highest level students use in most high school courses. 

Experiment Ideas:

If you decide to purchase a stereo microscope you can fill a small plastic tray with pond water and have them look through the side of the container with the microscope. If you decide to purchase a compound microscope you can purchase plain slides and cover slips and make your own slides out of vegetable peels, plants, and other transparent 3-D objects. 

Polarizing Microscopes Explained

A polarizing microscope is a microscope that is mainly used in geological studies to study geological specimens. For this reason, it is also known as a petrographic microscope. It is used in other scientific fields such as medicine and biology as well.

Polarizing microscopes are built like regular optical microscope, but are fitted with some extra features. Unlike regular microscopes which use normal light, a polarizing microscope uses polarized light to study specimens. In polarized light, the light waves vibrate in one direction; in normal light, the light waves vibrate in random directions.

Geological specimens ground into thin sections for study. The specimen to be studied is placed on a slide on a rotatable specimen stage. The specimen is then illuminated by a light source under the specimen stage.

We recommend our “Trinocular Infinity Polarizing Microscope with 1.3 MP Camera”

Product Features Include:

• Capable of bright field, and polarized light observation
• Infinity optical system, Kohler transmitted illumination
• 5 plan infinity strain free achromatic objectives
• Polarizer
• Analyzer
• Bertrand lens
• Compensator plates
• 1.3 MP digital camera