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Basic Malaria Microscopy (part I and II) (WHO; 1991; 72 pages)
Preface
Introduction
Learning Unit 1. Malaria, the condition
Learning Unit 2. Cleaning and storing microscope slides
Learning Unit 3. Maintaining accurate documents
Learning Unit 4. Blood films
Learning Unit 5. Staining blood movies with Giemsa stain
Learning Unit 6. The microscope
Learning Unit 7. Examining blood films
Learning Unit 8. Evaluating blood movies for malaria helminth
Learning Unit 9. Artefacts in blood movies
Learning Unit 10. Routine examination the blood films for malaria helminth
Learning Unit 11. Life bicycle of the malaria parasite
Learning Unit 12. Supervisory elements of malaria microscopy
Back sheathe

Learning Unit 6. The microscope


Learning objectives

By the finish of this Unit friend should:

• be able to demonstrate the usage of the microscopic lense with fabricated or herbal light• have the ability to demonstrate use of the oil immersion objective• have the ability to operate the mechanically stage• understand the surname of the main materials of the microscope• know just how to preserve the microscope and its components in great working order• understand what need to not be excellent to the microscope and also its components• know how to store the microscope• know exactly how to fill the microscope for transport from one ar to another.

You are watching: What can you infer about these two parts of a microscope?


You cannot perform your task without a microscope. It is necessary that friend learn just how to use it properly, the you know its limitations and that you know what demands to be done to store it in good condition.

The microscopic lense that friend will use is dubbed a compound microscope. Friend will require to recognize the surname of several of its component parts:

• so that you deserve to easily carry out instructions during the practical exercises the are component of your training

• so the you have the right to accurately explain parts that may need attention or replacement during the course of her work.

Parts that the link microscope

All the main parts that a common compound microscopic lense are illustrated in Fig. 1.

Main tube and also body tube

The key tube and also body tube space often jointly called the head the the microscope. The head typically slopes towards the user for greater comfort and also is then referred to as an skinny head. Polished glass prisms room fitted inside the body pipe of an lean head; these allow the light to bending so the the photo reaches the user’s eye.

The eyepiece, or ocular, is situated at the optimal of the key tube. Many compound microscopes are fitted through binocular heads, which way they have actually two oculars - one because that each eye. Some, however, have only one ocular and are described as monocular microscopes.

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Fig. 1 parts of a usual compound microscope

1. Key tube skinny head2. Body pipe (prism) skinny head3. Revolving nosepiece4. Objective5. Stage (mechanical stage)6. Substage condenser with iris diaphragm7. Mirror8. Basic (foot)9. Ocular (eyepiece)10. Eight (limb)11. Coarse adjustment12. Fine adjustment

Revolving nosepiece

A variety of objective lenses of different magnifications are screwed into the nosepiece that the microscope, which deserve to then it is in revolved to boost or decrease the magnification the the specimen gift examined.

Objectives

All components of the microscope are important, however the objective lenses - the lower, magnifying lenses - should be treated with particular care. The lenses room of the ideal quality and also need to be handled really carefully. Occasionally two lenses room glued together, and you have to be cautious not to use solvents together as solid alcohol options or acetone, which can dissolve the glue or cement.

Objectives are referred to by their magnifying power, i m sorry is significant on the side. The microscope you will use has the following objectives:

• x 10• x 40• x 100 (this objective is often referred to as the oil immersion objective; occasionally it has actually a black or red ring about it for basic identification).

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Objectives, reflecting magnification

You will notice in the diagram above that the dimension of the former lens the the target decreases together magnifying strength increases.

As the magnification differs in between objectives, therefore does the functioning distance. The functioning distance is the distance in between the prior lens the the objective and the specimen top top the phase (when the specimen is in focus). The higher the magnifying power of the objective, the shorter is the functioning distance. Working ranges for the standard missions are most likely to be as indicated below (depending on the make of the microscope):


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Objectives, showing working distance between front lens and also specimen

The mechanical stage

The mechanical phase holds the slide secure and permits the specimen come be relocated smoothly backwards, forwards or sideways. Sometimes a range is fitted to 2 sides that the phase to show the extent of the movement. This is dubbed the Vernier scale, and it is valuable to know how to usage it; it can be supplied to trace a component of the blood film the you have to re-examine or present to her supervisor.

Substage condenser (with iris diaphragm)

The substage condenser is made up of a number of lenses. These centre the irradiate from the mirror, or electric light source, to a central spot top top the field. The substage condenser deserve to be raised or lower to offer maximum or minimum illumination.

Inside the condenser is the iris diaphragm. This is used to regulate the lot of light passing through the condenser. The iris diaphragm is composed of a number of interlocking leaves made that a thin metal. The is changed by method of a lever.

Filter holder and also blue filter

Beneath the iris diaphragm is the filter holder. This is wherein a blue filter is placed when you use an electrical light resource for illumination. It has the effect of make the microscope field white quite than yellow.

Mirror

The mirror is supplied to straight light from the light resource to the microscope field. The mirror has actually two sides, one of which is a plane or level surface and is used with the substage condenser. The various other surface is concave and is used without the condenser (the curved surface itself acts together a condenser).

Note: some microscopes v a built-in illuminator execute not use a mirror however have a built-in prism instead, which directs light from the illuminator to the objective/ocular lens system. Others have actually a removable illuminator, which can be replaced by a mirror when necessary.

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Ocular (eyepiece)

The ocular or eyepiece fits into the upper end of the main tube and is what the microscopist looks through once using the microscope. The ocular has its magnifying power marked on it. Magnifying strength is the number of times through which it will certainly magnify the image produced by the objective. For instance, v an ocular the x 7 and an oil immersion objective of x 100, the full magnification the the specimen would be 7 x 100 = 700.

Oculars are accessible in a variety of powers. In malaria microscopy, an ocular of x 7 magnifying power is preferred. An ocular that x 6 could likewise be used, yet one of x 10 magnifying strength is not recommended.

Oculars equipment to binocular microscopes are dubbed paired oculars and are specially do to fit the microscope in question. On the pickled in salt of the oculars you may see the noting “ x 7P “, i m sorry denotes a paired collection of eyepieces the x 7 magnifying power.

Arm

The arm develops a rigid support for the main tube and also stage of the microscope. The is strong made and also can be used to carry the microscope roughly the laboratory. That is recommended, however, the you also support the microscope at the base, with your various other hand.

Coarse and fine adjustment

The two adjustment equipment - coarse and also fine - are offered to emphasis the specimen gift examined. The outlet adjustment is for rapid and also relatively huge movements of the stage (and therefore the specimen); the good adjustment is because that the finer concentrating required once the greater powered missions are used.

It is regular to emphasis the specimen an initial with the coarse adjustment and also then to use the well adjustment if the specimen is being examined.

With the oil immersion objective, the outlet adjustment is used in a different way. This will be described later.

Base (foot)

Whatever the shape of the basic of the microscopic lense (usually U-shaped or rectangular), it have to rest on a firm, level bench or table. That is important that the microscope does no wobble while the is gift used.

A threaded hole deserve to be checked out on the underside of the base. This is to take a screw the secures the microscope inside its storage box throughout transportation.

Use that the microscope

In the handy sessions you will certainly learn just how to use the microscope. You will certainly see exactly how the photo (of the specimen) shows up larger as you boost the magnification by changing objective lenses.

Initially friend will examine specimens provided to friend by her tutor or facilitator. Several of these will certainly be life organisms in water, others will be day-to-day objects with which friend are familiar (although they will certainly look an extremely different under the microscope).

During these exercises you will certainly learn how to adjust the illumination, and also see how to make the ideal use of the substage condenser and iris diaphragm. Girlfriend will likewise be able to use the mechanically stage and also the Vernier scale.

You will have time to practise ~ above a monocular microscope and will notice that its illumination is an extremely good. This is the microscopic lense that should be supplied with the x 100 oil immersion objective as soon as only organic light is available. The binocular microscope, though an ext restful because that long hrs of work, demands a reliable electrical energy supply because that illumination; through only organic light available, it is less effective than the monocular when provided with the oil immersion objective.

The irradiate source

A good source of irradiate is needed to research specimens properly. This may be either daylight or electric light. The electric current may be listed by mains it is provided or by a battery or generator. Light the is either as well bright or too dim will certainly interfere with examination of specimens.

Light native the light source travels the following path, via the substage mirror or the substage lamp:

light source → winter (if over there is one) → substage condenser and also diaphragm (if the source is artificial) → specimen → target → ocular

When artificial light is used, a blue filter must be placed in between the source and the substage condenser. If a mirror is work with synthetic light, the level side the the mirror should be used; once daylight is the light source, the concave mirror need to be used, there is no the substage condenser.

Obtaining also illumination

To obtain good, also illumination, the procedure is together follows:

Step 1 place the specimen slide on the mechanically stage. Utilizing the outlet adjustment, emphasis on the specimen with the x 10 objective.

Step 2 Making sure that the iris diaphragm is totally open, raise the substage condenser come the point where the field is brightest.

Step 3 eliminate the eyepiece and, while looking down the tube, change the mirror until the target lens is totally illuminated.

Step 4 replace the eyepiece and focus high solution on the specimen making use of the good adjustment.

Step 5 remove the eyepiece again and also close the iris diaphragm till the aperture that the target is two-thirds visible; this will certainly make the specimen appear clearer - it offers maximum resolution.

Step 6 replace the eyepiece and also revolve the nosepiece to pick the objective of the compelled power; you might need to emphasis slightly in ~ each readjust of objective. Certain directions worrying the use of the oil immersion target are offered below.

Illumination can be easily readjusted by enhancing or decreasing the aperture that the iris diaphragm.

Using the oil immersion objective

When setup up the microscope for use through the oil immersion objective, the following procedure is the best:

Step 1 after ~ arranging the illumination as explained in the vault section, watch the rest of the process from the next of the microscope.

Step 2 utilizing the outlet adjustment, rack up the key tube.

Step 3 place the on slide on the stage of the microscopic lense with the blood film uppermost.

Step 4 when you deserve to see the there will certainly be sufficient an are between the stage and the x 100 objective, turn the revolving nosepiece so that the x 100 target is end the specimen.

Step 5 place 1 - 2 fall of immersion oil ~ above the blood movie in the area i m sorry is to it is in examined.

Step 6 using the rough circuit adjustment, very closely lower the objective till the lens is in contact with the immersion oil. Climate raise the lens slightly, but enable the lens and oil to stay in contact.

Step 7 emphasis the specimen using the well adjustment, making certain that the lens does no come into call with the slide. Girlfriend may transform the illumination if necessary by adjusting the iris diaphragm.

Immersion oil is used in between the microscope slide and also the target lens to alleviate scattering the the light transmitted by the mirror or illuminator. The oil needs to reproduce the optical properties of the glass supplied for the lenses, and also must thus have a refractive table of contents of 1.515, i.e. Approximately 1.5 time the refractive table of contents of water.

When immersion oil is used, the target lens and also the slides must be cleaned in ~ the finish of the day’s work. You deserve to use a soft cotton cloth or lens tissue for the target lens yet remember no to usage it to clean various other lenses ~ above the microscope. From time to time, dried oil should be removed from the oil immersion objective making use of xylene (but no various other solvent). Oil deserve to be washed off the slides through a little amount that xylene; if no xylene is available, the oil smear deserve to be very closely dabbed with absorbent paper.

Immersion oil can be obtained commercially. In some countries, however, anisole is offered for job-related with the oil immersion objective; this product has actually the same refractive index as immersion oil. Anisole evaporates indigenous the blood film after some time, so the the film does not must be cleaned and there is much less chance the its gift damaged or delete everything off. Usage of anisole also method that the objective lens walk not must be cleaned.

Care of the microscope

Provided that normal care and common feeling are exercised, the laboratory microscope will be helpful for countless years.

Removing dust and grease

When not in use during the day, the microscope schould be retained covered v a clean fabric or plastic covering to protect the lenses from dust that settles out of the air. Overnight, or if the is to stay unused for long periods, the microscopic lense schould be placed inside that box v the door strictly closed. To safeguard the target lenses, the × 10 target schould be rotated to heat up through the ocular.

Oil and grease from eyelashes and fingers are quickly deposited ~ above lenses and oculars together the microscopic lense is used; these parts must be cleaned through lens organization or with very soft cotton cloth.

The oil immersion objective have to be cleaned after use. If that is no cleaned, the oil will certainly harden and also make the target useless. A lens organization or soft cotton cloth is usually adequate for the purpose. However, the tissue or towel should never be used to clean other objectives, the oculars or the mirror, otherwise oil will certainly be transferred to these components.

Preventing the development of fungus

In warm, humid climates it is really easy for fungal growths to come to be established on lenses and prisms. These growths can reason problems and may even become so poor that the microscope have the right to no much longer be used. The lenses may need to be repolished by the manufacturer, i beg your pardon is very expensive and may take numerous months.

Fungus cannot grow on glass as soon as the atmosphere is dry, and every initiative should therefore be made to keep the microscopic lense in a dry atmosphere when the is not being used. Among the following methods must be used:

• keep the microscopic lense in a repetitively air-conditioned room.

Note: that is meaningless to keep the microscopic lense overnight in a room where the air-conditioner operates only throughout the day.

• ar the microscopic lense in a “warm cupboard”, i.e. An airtight cupboard in i beg your pardon one or two 25-watt bulbs room constantly alight.

• store all lenses and prism heads in an airtight box or desiccator wherein the waiting is kept dry by method of active silica gel.

Note: Silica gelatin is a desiccant - a compound with the capability to absorb water vapour native the air. Self-indicating silica gel is blue when active but becomes pink when it has soaked up all the water it can. It deserve to then it is in reactivated through heating; it transforms blue again as it i do not care reactivated. As soon as the silica gelatin cools it have the right to be went back to the airtight container. Only self-indicating silica gel have to be used.

• In areas without electricity yet where a kerosene refrigerator is used, location the microscope box ~ above a small shelf sited 20 - 30 cm above the frozen refrigerator chimney. Warm from the chimney will store the crate sufficiently warm and also dry to stop fungus farming on the microscopic lense lenses.

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Transporting the microscope

When the microscope is to it is in transported indigenous one place to another, it is crucial to ensure the it is properly secure inside its box. The best means to perform this is by way of the securing device, i beg your pardon screws into the base of the microscope.