1. T20 TOCOPRODUCTS & Pvt. Ltd A COMPANY SERVE FOR STUDENTS

2. T20 EXAM PATTERN PROUDLY INTRODUCING BY T20 TOCO PRODUCTS & Ltd.

3. IN ASSOCIATION WITH THE TAMILNADU DR. MGR MEDICAL UNIVERSITY

4. REDUCING THE EXAM TIME BY ONE HOUR AND MARKS BY 50

5. I N T R O D U C E B R E A K A F T E R E A C H 3 0 M I N U T E S

6. G I V E F R E E H I T M A R K S F O R E V E R Y U N E X P E C T E D Q U E S T I O N S

7. FIRST 30 mins. POWER PLAY NO SUPERVISOR IN THE HALL

8. CHEER GIRLS IN EVERY CLASS. THEY WILL DANCE AFTER FININSHING EVERY PAGE

10. COUNTING OF BACTERIA

11. BACTERIAL COUNT COUNT OF LIVING AND DEAD CELLS TOTAL COUNT. COUNT OF ONLY LIVING OR VIABLE CELLS VIABLE COUNT.

12. TOTAL COUNT METHODS Direct methods: Direct methods: Breed’s method Breed’s method Wright’s method Wright’s method Counting Chamber Counting Chamber Coulter counter method Coulter counter method Membrane filtration Membrane filtration

13. Total count methods Indirect methods: Indirect methods: Visual method Visual method Photometric method Photometric method Colorimetry Colorimetry Nephlometry Nephlometry Packed cell volume Packed cell volume Dry weight Dry weight

14. VIABLE COUNT SOLID MEDIA TECHNIQUE SOLID MEDIA TECHNIQUE Pour plate method Pour plate method Surface viable method Surface viable method Drop technique Drop technique Spreading techinque Spreading techinque

15. VIABLE COUNT ROLL TUBE METHOD ROLL TUBE METHOD MEMBRANE FILTERATION MEMBRANE FILTERATION FLUID MEDIA TECHNIQUE FLUID MEDIA TECHNIQUE COUNTING ANEAROBES COUNTING ANEAROBES

16. TOTAL COUNT – BREED’S METHOD Microscopic slide – mark 01 sq.mm area Microscopic slide – mark 01 sq.mm area Place stage micrometer in the eye piece Place stage micrometer in the eye piece Dilute the suspension – spread carefully on the sq.mm area of the slide – dry it – fix it – stained – counted at the squares. Dilute the suspension – spread carefully on the sq.mm area of the slide – dry it – fix it – stained – counted at the squares.

17. DO YOU KNOW THESE? SIZE OF SQUARE IN THE EYEPIECE SIZE OF SQUARE IN THE EYEPIECE AREA OF FLIM ON THE SLIDE AREA OF FLIM ON THE SLIDE VOLUME OF SUSPENSION TAKEN on the slide VOLUME OF SUSPENSION TAKEN on the slide If you know ………………. If you know ………………. THEN THE NUMBER OF ORGANISMS IN THE ORIGINAL SUSPENSION CAN BE CALCULATED. THEN THE NUMBER OF ORGANISMS IN THE ORIGINAL SUSPENSION CAN BE CALCULATED.

18. WRIGHT’S METHOD Equal volumes of suspension and blood is intimately mixed. Equal volumes of suspension and blood is intimately mixed. Made a smear on the slide. Made a smear on the slide. Fixed – stained – counted Fixed – stained – counted Number of bacteria and red cells are counted for 40 fields. Number of bacteria and red cells are counted for 40 fields. Number of RBC - 5X10 6 per cm 2 Number of RBC - 5X10 6 per cm 2

19. As the ratio of blood cells to bacteria on the slide is known, the number of bacteria per cm 3 in the suspension can be found by proportionality. As the ratio of blood cells to bacteria on the slide is known, the number of bacteria per cm 3 in the suspension can be found by proportionality.

20. COUNTING CHAMBER Slide with recessed area that is ruled in squares. Slide with recessed area that is ruled in squares. Drop of culture is placed over that and place a cover slip. Drop of culture is placed over that and place a cover slip. Don’t allow any air to entrap Don’t allow any air to entrap Allow to settle the cells on the area Allow to settle the cells on the area View under phase contrast microscope View under phase contrast microscope Stain and focus under normal micros. Stain and focus under normal micros.

21. Number of organisms in 04 groups of 16 small squares are counted Number of organisms in 04 groups of 16 small squares are counted CIRCULAR DITCH CELLSTRAIGHT DITCHES Cover glass Ditches Grid Depressed cell Suspension cel End view

23. COULTER COUNTER METHOD Flow of suspension Tube Electrode Orifice Suspension

25. MEMBRANE FILTRATION Provided that the suspension could be sufficiently diluted Provided that the suspension could be sufficiently diluted Organism retained on the filter are stained – dried – counted under microscope Organism retained on the filter are stained – dried – counted under microscope The number of bacteria on the filter paper, from a known volume of suspension can be calculated The number of bacteria on the filter paper, from a known volume of suspension can be calculated Mostly applicable when the numb of organism is very small.

26. INDIRECT METHODS COMPARISION WITH STANDARD SUSPENSION

27. Use of std. opacity tubes. Use of std. opacity tubes. 10 tubes – solid suspn. of unglazed porcelain in increasing OD. 10 tubes – solid suspn. of unglazed porcelain in increasing OD. They are standardized against accurately counted bacterial Suspension. They are standardized against accurately counted bacterial Suspension. Comparing the opacity with standards. Comparing the opacity with standards. VISUAL METHOD

28. PHOTOMETRIC METHOD CALORIMETRY CALORIMETRY NO. OF CELLS METER READING LIGHT SOURCE SPECIAL FILTER SHUTTER OPTICAL CELL PHOTO CELL AMOUNT OF LIGHT ABSORBED BY THE SUSPENSION IS CALCULATED

29. NEPHLOMETRY THE AMOUNT OF LIGHT REFRACTED BY THE PARTICLES IN THE SUSPENSION IS MEASURED

30. PACKED CELL VOLUME Suspension is centrifuged Suspension is centrifuged Depth of sediment is measured Depth of sediment is measured Equivalent num. of cells is obtained from a graph of cell numbers against paced cell volume Equivalent num. of cells is obtained from a graph of cell numbers against paced cell volume

31. DRY WEIGHT Centrifuged cells – washed, Centrifuged cells – washed, dried – weighed Weight can be related to number of cell Weight can be related to number of cell

32. VIABLE COUNT ROLL TUBE METHOD SOLIDIFIED AGAR MEDIA LAYER COLONIES The rubber closure should be loosened during incubation

33. MEMBARANE FILTRATION MEMBRANE FILTER IMPREGNATED WITH MEDIUM BY STEREOSPECIFIC MICROSCOPE AND SURFACE ILLUMINATOR – COUNTING CAN BE DONE USING A FILTER OF 47 TO 50 MM DIAMETER – UP TO 200 COLONIES CAN BE COUNTED

34. FLUID MEDIA TEECHNIQUE 1 – 1 0 t e s t s u s p e n s i o n d i l u t i o n i n s e r i a l – f i n d t h e t u r b i d i t y – w i t h d i l u t i o n f a c t o r – c o u n t i n g i s d o n e

35. THANK YOU I ACKNOWLEDGE MY MISTAKES IF ANY Meet again