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Microscopes

How does a microscope work?

A microscope works by producing an enlarged image of a small object.

Because the eye can only focus to about 10 inches (25cm), this limits its ability to see fine detail by simply moving in closer. A magnifier has a much shorter focal length and produces a beam of parallel rays that the eye treats as coming from infinity. The eye has no problem focusing these rays, so it can now comfortably see a blown-up image as if it were at the position of the magnifying glass. In essence, the magnifier lets the eye work at a much closer distance where it can see a larger image.

A microscope works the same way as a simple one-element magnifier, using a series of lenses to provide a very short effective focal length and correspondingly high magnifications.


What do the numbers on the barrel of the microscope objective mean? What about the letters DIN and JIS?

Microscope objective lenses will often have 4 numbers engraved on the barrel in a 2x2 array.

The upper left number is the magnification factor of the objective. For example, 4x, 10x, 40x 100x.

The upper right number is the numerical aperture of the objective. For example 0.1, 0.25, 0.65 and 1.25.

The lower left number is the tube length in millimeters. This is related standardization of microscopes and the particular standard used for the manufacture of your microscope. Most microscopes employ the Deutsche Industrie Norm, or DIN standard configuration. The Japanese Industrial Standard (JIS) is less commonly used. DIN microscopes begin with an object-to-image distance of 195mm, and then fix the object distance at 45mm. The remaining 150mm distance to the eyepiece field lens sets the internal real image position, which is defined as 10mm from the end of the mechanical tube (which gives the 160mm tube length). DIN standard eyepieces have an international standard 23mm diameter. DIN standard objectives often times have "DIN" etched on the side and have a standard 0.7965" diameter thread, 36 TPI, 55° Whitworth threading.  Celestron microscopes are made to DIN standards. The tube length for the DIN standard is 160mm, while for the JIS it is 170mm.

NOTE: JIS objectives can be used on a DIN microscope and vice versa  The threads on both types are interchangable. However, since the optical distances are different, there will be a difference in magnification.

The lower right number (if given) refers to the thickness of the glass cover slip (in millimeters) assumed by the lens designer for best performance of the objective. Example: 0.17.

Sometimes objectives have a color ring to aid in identifying the magnification: black (1x), brown (2x), red (4x), yellow (10x), green (20x), turquoise (25x), light blue (40x), dark blue (60x), white (100x).
Magnification: how do I calculate it for my microscope?

It’s very easy to figure out the magnification of your microscope. Simply multiply the magnification of the eyepiece by the magnification of the objective lens.

The magnification of both microscope eyepieces and objectives is almost always engraved on the barrel (objective) or top (eyepiece). Look for numbers like 10x, 12.5x, etc.
My microscope has a filter wheel. How is it used?

Filters are used to increase contrast and color correction for visual observations of specimens or slides. They may be used with live specimens as a non-lethal substitute for staining.

Blue is commonly used to correct yellowish or reddish color from incandescent bulbs in illuminators. Other filter colors (green, yellow, magenta, frosted) should be tried with specimens to see if they improve contrast and visibility. Simply turn the wheel to put the filter you need into the path of light from your condenser-illuminator.
What is the difference between achromatic, semi-plan and plan objectives?

Most microscope objectives come in three basic designs: achromatic, semi-plan and plan.

An achromatic objective corrects for color and has a flat field across the central 65% of the image. It’s usually a pair of lenses.

Semi-plan (semi-planar, sometimes called micro-plan) objectives have an 80% flat field. They can either be two lens element achromats or three or more element apochromats.

A plan (or planar) objective corrects better for color and spherical aberration than either the semi-plan or the achromatic objective. Plan objectives have a flat field about the center 95% of the image. They also often have larger working distances. While plan objectives give you flatter fields than achromatic objectives, they also are the most expensive.
What’s the resolution of a microscope? Is this related to numerical aperture?

A microscope’s resolution is its ability to form separate images of lines or dots. It’s defined in terms of the actual distance between the detail on the object. The wave nature of light puts a practical and theoretical limit on the resolution of a white-light microscope as about half a wavelength of visible light, about 250 nanometers or a quarter-micron. This also limits the highest useful magnifications to the range of 1000x-2000x. Resolution depends solely on the objective lens, as the eyepiece just magnifies light from the objective.

The numerical aperture of a microscope lens is a measure of the increase in resolution due to the lens being able to gather a wider cone of light from the specimen. This depends on how close the lens is to the specimen and also if oil or another fluid is used between the lens and the specimen.  Both increase the numerical aperture and the microscope’s resolving power.

Numerical aperture is higher for higher-power objectives because they are used very close to the specimen. Its value for the objective is usually engraved on the barrel and runs between 0.04 for very low-power objectives and 1.4 for very high-power objectives. A typical 40x objective that will be used at 400x with a typical 10x eyepiece has a numerical aperture of 0.65.

A condenser’s numerical aperture should equal or exceed that of the objective lens to effectively illuminate the specimen. Higher powers will need condensers with higher numerical apertures, meaning the condenser lens is closer to the specimen. Condensers with numerical apertures of 1.2 are typical for objectives delivering magnifications of 400x.
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Which are the main features of a microscope?

Before buying a microscope, it's important to keep in mind the following characteristics:
- MAGNIFICATION: the product of eyepiece magnification and lens magnification gives the total magnification power. An increased magnification decreases the field of vision.
- ILLUMINATION: it may be either incident light, meaning from below and particularly useful in a biological microscope, or transmitted light, meaning from above and for a stereoscopic microscope. Some stereoscopic models have both types of illumination.
 
Which are the optional accessories recommended for a microscope?

According to client's needs, in order to increase or decrease the magnification provided, it could be necessary to purchase extra eyepieces and/or lenses.
In a biological microscope, when it isn't included, a small transport table (a square-shaped instrument that allows microscopic movements of the glass plate) and a light to provide the necessary lighting in the observation of the prepared slides, could be useful.
A special graduated micrometric eyepiece could also be useful, in order to measure exactly the dimensions of the prepared slides
 
Can a camera be connected to a microscope?

Yes, but two adapters are necessary: the photo adapter for a given microscope and the T-2 ring for the type of reflex. On a binocular microscope the camera is connected to one of the two eyepieces.
 
Is it always necessary to prepare the items for the observation at the microscope ?  

Using a stereoscopic microscope, whatever is viewed doesn't need preparation. Instead, using a biological microscope, it's necessary to make accurate preparations. The sample must be cut very thin and it must be placed on a glass slide. It could seem a rather complicated process, so if you don't want to do it yourself, Konus has in stock more than 20 series of prepared slides with interesting biological subjects.
 

Information on this page courtesy Celestron’s web site celestron.com
For all international order please email for S&H fees - Celestron Products can only be shipped within the USA
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