Laboratory Training. Summer Internship 2015 ACRM

Transcription

1 Laboratory Training Summer Internship 2015 ACRM

2 Pipetting Volumes: 1000µL, 100µL, 10µL 15 repeats each Acceptable variation 5% Method Pipette distilled H20 onto a balance and record the weight

3 Microbeads Concentrations: High, Low 3 repeats each Acceptable variation 15% Method Fill microcell and perform count in 4 fields Calculate average and concentration Validate against reference values

4 Sperm Concentration Evaluate 10 Semen samples Acceptable variation 20% Method Fill microcell and perform count in 4 fields Calculate average and concentration Validate and compare to mentor s results

5 Sperm Motility Evaluate 10 Semen samples Acceptable variation 20% Method Fill microcell and perform count in 4 fields Calculate average and % motility Validate and compare to mentor s results

6 Sperm Vitality Prepare 2 different smears Evaluate 5 reference slides Acceptable variation 15% Method Evaluate each slide x 2 Count viable and non-viable cells (100 sperm) Calculate average and concentration Validate and compare to reference values

7 Data Recording Record all information in your lab book Transfer to day sheet Populate soft copy (Excel) Print and have it verified

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9 Summer Internship Bench Research Training The interns will observe and practice the following techniques prior to the actual start of their original research projects. Laboratory safety: Explain the dress code that needs to be observed (lab coats, closed shoes, etc.), designated work areas, and the basic safety procedures to be followed. Pipetting: Explain the different types of pipettes (transfer pipette, micropipette, pipettor), when they should be used and how they should be operated. Exercise - Pipette 3 volumes (1000µL, 100µL and 10µL) x 15 repeats Microscopy: Explain the various types of microscopes, the function of different components as well as the proper operating and cleaning procedures (Tutorial) General handling of a semen sample: Explain and demonstrate the basics to be performed when a semen sample was received: Incubation, Recording of information, Liquifaction, transfer, volume, ph, Viscosity, Color Microbeads: Explain the loading of a microcell, setting up of the microscope (x20 phase contrast), counting technique and calculations Exercise Count 2 concentrations x 3 each Sperm count and Concentration: Vitality Explain the loading of a microcell, setting up of the microscope (x20 phase contrast), counting technique and calculations Exercise Analyse 10 x samples Explain the principles of employing a dye exclusion staining technique for viability measurement, preparing the sample, making of the smear, coverslip slide, and evaluating the sample Exercise Prepare and score x 5 slides Morphology: Explain the principles of morphology, the preparation, staining and evaluating of slides Demonstrate Slide preparation, staining, scoring Exercise Only identified students will receive further training

10 TUNEL ROS ORP Explain the principles of TUNEL as well as the calculations and preparation method. Exercise Only identified student will be receive further training Explain the principles of ROS measurement Demonstrate Perform the sample preparation, machine setup and calculation Exercise Only identified student will be receive further training Explain the principle of sorp and corp Demonstrate Perform sample preparation, machine setup and calculation Specialized Techniques Techniques specific to projects (e.g. Acrosin activity, Western Blot analysis, Proteomics, HOS, etc.) will be explained by the particular mentors to the individual(s) responsible in each group

11 Summer Internship 2015 Bench Research Training Proficiency Report Intern: Mentor:

12 Pipetting Proficiency Pipette: P-1000 Reading Actual volume (µl) Average Coefficient of Variance (CV) Measured volume (µg) Absolute Percentage Acceptable variation: 5% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

13 Pipette: P-200 Reading Actual volume (µl) Average Coefficient of Variance (CV) Measured volume (µg) Absolute Percentage Acceptable variation: 5% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

14 Pipette: P-200 Reading Actual volume (µl) Average Coefficient of Variance (CV) Measured volume (µg) Absolute Percentage Acceptable variation: 5% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

15 Counting Microbeads: Low concentration Reading Reference Count (x10 6 /ml) Average Coefficient of Variance (CV) Measured Count (x10 6 /ml) Absolute Percentage High concentration Reading Reference Count (x10 6 /ml) Average Coefficient of Variance (CV) Measured Count (x10 6 /ml) Absolute Percentage Acceptable variation: 15% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

16 Sperm concentration: Reading Average Reference Count (x10 6 /ml) Coefficient of Variance (CV) Measured Count (x10 6 /ml) Absolute Percentage Acceptable variation: 20% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

17 Motility: Reading Average Reference Motility Coefficient of Variance (CV) Measured Motility Absolute Percentage Acceptable variation: 20% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)

18 Vitality (E&N): Reading Average Reference Viability Coefficient of Variance (CV) Measured Viability Absolute Percentage Acceptable variation: 15% Met the criteria: Yes / No Retraining completed: Not Applicable / Yes / No Verified: (Mentor) Approved: (Director)