Standard Operating Procedure of Operation, Cleaning and Calibration of Microscope.

1.0 Purpose:
To lay down the procedure for operation and calibration of microscope.

2.0 Scope:
This procedure is applicable for operation and calibration of microscope in Microbiology Laboratory.

SOP for Operation, Cleaning and Calibration of Microscope

3.0 Responsibility:
Microbiologist / Designee :
•To perform operation procedure as per procedure.

•To perform calibration as per calibration procedure.

•To fill calibration data as per format.

Head/Designee - QC :
•To review, implement and ensure compliance of SOP.

Head/Designee - QA :
•To approve, implement and ensure compliance of SOP.

4.0 Procedure:

4.1 Precaution:

4.1.1 Handle the microscope carefully.

4.1.2 Do not leave dust, dirt or finger marks on the lens surfaces.

4.1.3 Clean the dust using soft brush, finger marks with soft cotton cloth. Use xylene for cleaning the objectives & immersion oil. Avoid organic solvents.

4.1.4 If bubbles are trapped between the objective lens & the oil on the slide, start again. Keep the oil tightly stoppard & do not shake the bottle.

4.2 Operation:

4.2.1 Make sure all the exposed optical surfaces are free from dirt.

4.2.2 Plug the power cord and bring the light intensity regulator to lowest level & put “ON” the power.

4.2.3 Turn “ON” the main switch and power switch of the microscope.

4.2.4 Adjust the diaphragm and intensity controller to get sufficient light towards the stage micrometer.

4.2.5 Place the material/specimen to be tested on a slide and spread it uniformly with the help of another slide and cover it with a cover slip (except for oil immersion specimens).

4.2.6 Set the magnification as per requirement.

4.2.7 Select a suitable objective by rotating the objective holder and adjust the specimen to bring it into focus by means of the coarse & fine adjustment knob, while looking through the eyepiece.

4.2.8 For a microbial specimen slide, place a drop of immersion oil on the smear, adjust the objective by touching it to the oil and observe through the eye piece to bring the specimen into focus by using the fine adjustment knob.

4.2.9 The Trinocular port has a CCD camera connected. This helps us to view the image as seen in the ocular eyepiece on a computer monitor and also store the same on PC.

4.2.10 On the right hand side near the viewing head, there is a knob to select the image to be either seen by the eyepiece or to be transferred to the CCD camera for the future transfer to PC.

4.2.12 For viewing in the PC, push the knob and for viewing by eye pull the knob.

4.2.13 Switch “ON” the camera.

4.2.14 In the software on clicking application icon, live image shall be displayed on PC screen.

4.2.15 After select the proper field of view the image can be stored directly to the PC.

4.2.16 After completing observation shut down the program and PC and also switch “OFF” the camera and microscope.

4.3 Calibration:

4.3.1 Place the Ocular micrometer in the eye piece.

4.3.2 Place the stage micrometer on the stage.

4.3.3 Focus with the help of the coarse adjustment knob and if required adjust with the help of fine adjustment knob.

4.3.4 One division of the stage micrometer is equal to 0.01 mm.

4.3.5 Check the no. of divisions coinciding of stage micrometer to that of ocular micrometer at 10 x 40X and 10 x 100X magnifications.

4.3.6 Calculate the division equivalence in µm for each magnification by formula.

                      No. of division of stage micrometer
                 = ------------------------------------------------- * 10
                      No. of division of ocular micrometer

4.3.7 Record the data in the calibration format as per format.

4.3.8 Calibration Frequency:

4.3.8.1 Once in a six months ±15 days.

4.4 Maintenance:

4.4.1 When instrument is under breakdown put the tag stating “UNDER MAINTENANCE” with date and sign of the analyst as per format and shall inform to QC Head/designee.

4.4.2 QC Head/designee shall inform to the engineering person or instrument manufacturer / service person.

4.4.3 After completion of major repairing / servicing job, calibrate the instrument to ensure the proper working. Maintain the record as per respective calibration format.

4.4.4 After servicing of the instrument attach copy of service report prepared by the instrument engineer.

4.5 Cleaning:

4.5.1 Clean the instrument with a clean, soft, dry cloth every day, taking care that the lenses are not scratched.