semen analysis and preparation dr. sushma ved consultant embryologist
DESCRIPTION
SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST. SEMEN ASSESMENT SELECTION OF AN APPROPRIATE METHOD OF PREPARATION. PURPOSE OF PREPARATION. YIELD OF HIGH PERCENTAGE OF FUNCTIONAL PROGRESSIVELY MOTILE - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/1.jpg)
SEMEN ANALYSIS AND
PREPARATION
DR. SUSHMA VED
CONSULTANT EMBRYOLOGIST
![Page 2: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/2.jpg)
SEMEN ASSESMENT SELECTION OF AN APPROPRIATE
METHOD OF PREPARATION
![Page 3: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/3.jpg)
PURPOSE OF PREPARATION
YIELD OF HIGH PERCENTAGE OF
FUNCTIONAL
PROGRESSIVELY MOTILE
MORPHOLOGICALLY NORMAL SPERMATOZOA
ELIMINATE SEMINAL PLASMA – CONATINS CYTOKINES AND PROSTAGLANDINS
![Page 4: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/4.jpg)
EJACULATED SEMEN IS VISCOUS FLUID AND MIXTURE OF
SEMINAL PLASMA
TESTICULAR AND EPIDIDYMAL SECRETIONS
POLYGONAL EPITHELIAL CELLS AND LEUCOCYTES
LIVE AND DEAD SPERMS
![Page 5: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/5.jpg)
SEMEN COLLECTION
3-5 DAYS ABSTINENCE
60-100 ml WIDE MOUTH STERILE CONTAINER
WITH H/O PSHYCOLOGICAL STRESS FACTOR
• COLLECTED AT HOME• TRANSPORTED TO HOSPITAL AT BODY
TEMPERATURE , 30-60 min
![Page 6: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/6.jpg)
![Page 7: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/7.jpg)
CRYOPRESERVATION – IF NEEDED
VISCOUS, REDUCED VOLUME 2-3 ml OF CULTURE MEDIUM ADDED
![Page 8: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/8.jpg)
EJACULATORY DYSFUNCTION
RETROGATE EJACULATION
OBJECTIVE: pH - 7.4 OSMOLARITY- 320 mosm/kg
![Page 9: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/9.jpg)
1. ORAL INTAKE OF SODIUM BICARBONATE AND WATER
2. ALTERNATIVELY – CULTURE MEDIUM INJECTED INTO THE BLADDER BEFORE EJACULATION
3. CENTRIFUGE IMMEDIATELY
![Page 10: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/10.jpg)
ANEJACULATION
PENILE VIBRATORY STIMULATION
ELECTROEJACULATION
![Page 11: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/11.jpg)
LIQUIFICATION
•ALLOW TO LIQUIFY NATURALLY - 30-45 mins AT 37⁰C •WHETHER SAMPLE RUNS FREELY ON PIPETTING•VISCOUS SAMPLES ARE DIFFICULT TO PIPETTE•REPEATED ASPIRATION THROUGH THE PIPETTE(with the addition of culture medium)HELPS TO BREAK VISCOSITY
![Page 12: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/12.jpg)
![Page 13: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/13.jpg)
NOTE AND RECORD THE COLOUR
MIX SEMEN THOUROUGHLY
MEASURE THE VOLUME
![Page 14: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/14.jpg)
SEMINAL ANALYSIS WHO minimum standard,2002
VOLUME 1.5 ml or more PH >7.2 DENSITY 20m/mlTOTAL SPERM NO 40 m/ejaculateMOTILITY > 50% PROGRESSIVE OR
>25% RAPIDLY PROGRESSIVE
MORPHOLOGY > 30% NORMAL FORMSWHITE BLOOD CELLS < 1 million/ml
![Page 15: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/15.jpg)
COUNT AND MOTILITY
FIX DEPTH MAKLER CHAMBER IS USED
COUNT AND MOTILITY IS ASSESED SIMULTANEOUSLY
![Page 16: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/16.jpg)
MAKLER CHAMBER
![Page 17: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/17.jpg)
MAKLER CHAMBER
![Page 18: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/18.jpg)
![Page 19: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/19.jpg)
COUNT / ml - TOTAL NUMBER OF SPERMS IN 10 SQUARES
% MOTILITY =(No. OF MOTILE SPERMS IN 10 SQUARES OF CHAMBER)X100 / TOTAL NO. OF SPERMS IN 10 SQUARES
![Page 20: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/20.jpg)
![Page 21: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/21.jpg)
MOTILITY ACCORDING TO WHO Class a: Rapid,progressive –linear
forward progression – 25 microm / sec
Class b: Slow progressive-5 -25 microm /sec
Class c: Non progressive motility(flagellar activity) .
Class d: Immotile (no flagellar activity)
![Page 22: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/22.jpg)
MORPHOLOGYTYGERBERG STRICT CRITERIA-14%
HEAD SHAPE/SIZE DEFECTS
NECK/MIDPIECE DEFECTS
TAIL DEFECTS
![Page 23: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/23.jpg)
![Page 24: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/24.jpg)
![Page 25: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/25.jpg)
![Page 26: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/26.jpg)
RELATED TO INFERTILITY-
1. GLOBOZOOSPERMIA 2. IMMOTILE CILIA SYNDROME-
KARTAGENER SYNDROME
![Page 27: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/27.jpg)
MEDIA SPERM PREP MEDIA
SPERM RINSE – VITROLIFE(SWEDEN)
GAMETEBUFFER – COOK(AUSTRALIA)
SPERMPREPMEDIA - MEDICULT
![Page 28: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/28.jpg)
BUOYANT DENSITY GRADIENT
PURE SPERM (SCANDINAVIAN IVF)
IXTA PREP (MEDICULT)
SIL SELECT (MICROM)
ISOLATE(IRVINE SCIENTIFIC)
![Page 29: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/29.jpg)
METHODS OF PREPARATION
1. CENTRIFUGATION – SWIM UP
2. SWIM UP FROM SEMEN
3. BUOYANT DENSITY GRADIENT
4. GLASS WOOL FILTRATION
STERILE TECHNIQUE MUST BE FOLLOWED THROUGHOUT
![Page 30: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/30.jpg)
CHOICE OF PREPARATION
DEPENDS ON- MOTILE COUNTRATIO BETWEEN MOTILE AND IMMOTILEVOLUMEVISCOSITYPRESENCE OF ANTIBODIES,AGLUTINATION,PUS CELLS AND DEBRIS
![Page 31: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/31.jpg)
DIRECT SWIM UPMOST OF THE SAMPLES CAN BE PROCESSED EXCEPT - WITH LOW MOTILITY AND POOR PROGRESSION
1-1.5 ml OF CULTURE MEDIUM TAKEN IN TWO ROUND BOTTOM TUBES(MULTIPLE TUBES CAN BE USED)
GENTLY PIPETTE 1ml OF LIQUIFIED SEMEN UNDERNEATH MEDIUM
![Page 32: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/32.jpg)
![Page 33: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/33.jpg)
![Page 34: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/34.jpg)
ALLOW TO STAND FOR 30-60 mins AT 37⁰C- SPERM MIGRATING TO OVERLAID MEDIUM-TURNS OPAQUE OR MILKY
CAREFULLY ASPIRATE ALL THE SUPERNATANT.
CENTRIFUGE COLLECTIVELY IN 15ml CONICAL TUBE AT 1500 – 2000 rpm FOR 10 mins
![Page 35: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/35.jpg)
![Page 36: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/36.jpg)
![Page 37: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/37.jpg)
![Page 38: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/38.jpg)
![Page 39: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/39.jpg)
ASPIRATE THE SUPERNATANT – VERY CAREFULLY WITHOUT DISTURBING THE PELLET
AT 0.3 – 0.5ml OF IVF MEDIUM TO THE PELLET AND MIX
LOOK FOR COUNT AND MOTILITY
![Page 40: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/40.jpg)
![Page 41: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/41.jpg)
![Page 42: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/42.jpg)
![Page 43: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/43.jpg)
DIRECT SWIM UP
ADVANTAGE - CENTRIFUGATING ONLY MOTILE SPERMS WITHOUT LEUCOCYTES,DEAD SPERMS AND DEBRIS IN THIS WAY REDUCING THE EFFECT OF ROS
CENTRIFUGATING ONLY ONCE MINIMIZING THE TRAUMA OF CENTRIFUGATION
![Page 44: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/44.jpg)
BUOYANT DENSITY GRADIENT PREPARED FOR SAMPLES WITH LOW MOTILITY AND POOR PROGRESSION
1. A TWO LAYERED GRADIENT COLUMN IS PREPARED BY PLACING 1ml OF 90% SOLUTION AT THE BOTTOM OF CONICAL TUBE AND ADDING 1ml OF 45% SOLUTION OVER THE 90%
2. 2 ml OF LIQUIFIED EJACULATE IS LAYERED OVER THE GRADIENT
3. CENTRIFUGE FOR 10mins AT 2000 rpm
![Page 45: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/45.jpg)
![Page 46: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/46.jpg)
![Page 47: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/47.jpg)
![Page 48: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/48.jpg)
4. REMOVE THE SUPERNATANT , ADD 2 ml OF CULTURE MEDIUM TO THE PELLET AND MIX
5. CENTRIFUGE FOR 10 mins
6. REMOVE THE SUPERNATANT
7.MIX THE PELLET WITH 0.3 – 0.5 ml OF CULTURE MEDIUM
![Page 49: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/49.jpg)
![Page 50: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/50.jpg)
![Page 51: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/51.jpg)
![Page 52: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/52.jpg)
Semen parameters vary from individual to individual and within individual too.
The sample given on the day of procedure can be entirely different from previous assesment
![Page 53: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/53.jpg)
MOST OF THE SAMPLES CAN BE PREPARED BY DIRECT SWIM UP EXCEPT SEMEN WITH LOW MOTILITY AND POOR PROGRESSION WHERE BUYONT DENSITY GRADIENT METHOD IS PREFERRED.
![Page 54: SEMEN ANALYSIS AND PREPARATION DR. SUSHMA VED CONSULTANT EMBRYOLOGIST](https://reader035.vdocuments.net/reader035/viewer/2022081422/5681661f550346895dd97244/html5/thumbnails/54.jpg)
THANK YOU