Calibration of filar eyepiece micrometer
A feature of the use of filar and stage micrometers for determining the rate and scale in biological cells and tissues. The characteristic of measuring the size of cells in a microscopic slide using the universal instrument of the ophthalmic eyepiece.
| Рубрика | Иностранные языки и языкознание |
| Вид | лабораторная работа |
| Язык | английский |
| Дата добавления | 16.07.2017 |
| Размер файла | 11,4 K |
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LUBLIN UNIVERSITY OF TECHNOLOGY
ENVIRONMENTAL BIOLOGY
FUNDAMENTALS OF MICROSCOPY
Lublin 2015
Aim of Classes
Focusing on microscope technique and using filar and stage micrometers to determine size and scale in biological cells and tissues.
Apparatus and Materials
Microscope , filar eyepiece micrometer,stage micrometer , sharp knife or scalpel, small onion , slides and cover slips , forceps, mounted needle , tweezers, dropping pipette, 0,85%NaCl solution
1. Preparing a slide of onion epidermal cells
1. Cleaning a slide
2. Placing a drop of 0,85% NaCl solution in the center of the slide.
3. Cutting open an onion and separating some of the leaves, peeling off one of the thin layers of epidermal tissues from the inner concave surface of a leaf.
4. Transferring sample to a drop of 0,85 % NaCl solution on a microscope slide - using tweezers and a mounted needle to making sure that the tissue is not folded. filar micrometer tissue ophthalmic
5. At an angle , placing one slide of the cover slip against the slide making contact with outer edge of the liquid drop, gently lowering a cover slip onto the slide , avoiding air bubbles.
6. Placing the slide on the stage of the microscope and using the low-power objective to locate the cells. After using the high-power objective to select several cells that are clearly visible in your field of view.
7. Making a large , labelled drawing of several epidermal cells . Making drawings of several epidermal cells at magnification of objective lens : 5x, 10x,40x.
2. Calibration of filar eyepiece micrometer
1. Placing the stage micrometer on the microscope stage and focusing as usual until the lines appear sharp .
2. Using the mechanical stage , center the stage scale in the field of view so that the vertical lines are oriented North-South and the stage scale centered in the field of view from "9 o'clock to 3 o'clock positions .
3. Removing the visual eyepiece from the microscope body tube .
4. Holding the filar eyepiece toward a bright light . Using the adjustable eye lens of the eyepiece , focusing the field of view fixed scale (0-8) and X hairs so that they appear sharp .
5. Inserting the filar eyepiece into the microscope tube and focusing until the lines of stage microscope appear sharp .
6. Setting the fixed scale of the filar eyepiece micrometer parallel to the scale of the stage micrometer .
7. Without changing the adjustment of the eye lens of the filar micrometer ,tightening the retaining screw of the filar eyepiece so that it is fixed to the microscope body tube . The scale in the field of view will be seen and if the drum is turned so that the fiduciary pair of lines is at 4 in the field of view and the external drum reading is at 0 , the X hairs will be seen in the center of the field of view .
8. Using the mechanical stage , moving the stage micrometer scale so that a longer vertical line ( a tenth division ) is exactly intersected by the traveling X hair of the filar.
9. Starting with the external drum at 0 and the X hair intersecting the longer vertical line of the stage scale , rotating the external drum toward the user (going from 0 upwards ) . While looking at the X hairs , moving the X hairs so that the X hairs and fiduciary lines travel a distance of 10 small spaces on the stage scale . Recording the "starting" number on the fixed scale and the "finishing" number on the fixed scale; also recording the number on the external drum .
10. Using the data obtained performing the necessary calculations.
11. Calibrating the filar eyepiece micrometer at magnification of objective lens : 5x , 10x , 40x .
3. Measurement of cells size in microscopic slide using filar eyepiece micrometer
1. Placing the microscope slide on the stage and faster it with the stage clips.
2. Removing the visual eyepiece from the microscope body tube and inserting the filar eyepiece into the microscope tube .
3. Focusing on the object to be measured at fixed magnification of objective lens.
4. Starting with the external drum at 0 and X hair intersecting the edge of the measured object , rotating the external drum toward the user ( going from 0 upwards ).
5. While looking at the X hairs, moving the X hairs so that the X hairs and fiduciary lines travel a distance of measured section .
6. Recording the "starting" number on the fixed scale and the "finishing" number on the fixed scale; also recording the number on the external drum .
7. Recalculating the total small drum divisions according to calibration of filar eyepiece micrometer for the user magnification of objective lens.
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