Supplies, equipment and methods

Methods

1. Collect semen sample in a sterile cup.
2. Wait for semen to liquify at room temperature; take HardyCHROM Candida out of the refrigerator, and warm to room temperature (1 hour).
3. Use a vortex mixer to ensure uniform suspension of cells in semen.
4. Using a pipette, collect 2 ul of semen and drop on culture medium. To detach the 2 ul drop from the pipette tip, slowly lower the drop until it touches the agar surface. Do not touch the medium with the pipette tip. Avoid placing drops close to the edge of the Petri dish or on printed sections, as this will prevent visualizing growth using a microscope. Place drops 15 mm apart from each other, allowing up to 16 drops to be placed per dish in a 4 x 4 matrix.
5. Wait for the drops to be absorbed by the culture medium (15 minutes).
6. Incubate up-side-down at 31C for 24 hours or more.
7. Delicately place a cover glass on agar surface. Avoid placing the cover glass close to the edge of the Petri dish, as this will prevent it from being flat on surface.
8. Place Petri dish on microscope stage; place a drop of high viscosity microscope oil on the cover glass; view results with 10x and 100x lenses (for a total magnification of 100x and 1000x).

Supplies and equipment

 Supply / equipment
 Supplier Cost
 Microscope (x2000 w/ 1.3M digital camera) Amscope 230
 Microscope cover glass 24 x 24 mm No. 1
 Globe Scientific
 25
4 oz Specimen Container, Red Screwcap, Sterile
 Globe Scientific  22
 2 - 20 uL Diamond Pipettor
 Globe Scientific  150
 0.1 - 10µL Pipette Tip, Racked, STERILE Globe Scientific 48
 Incubator (ex: Exoterra by Hagen)
 Hagen 120
 HardyCHROM Candida
 Hardy Diagnostics
 25
  620

Other suppliers
www.p212121.com (for chromogenic substrates).

Comments