Measuring mouse sperm parameters using a particle counter and sperm quality analyzer: a simple and inexpensive method.
This study examined a method for analyzing the count, motility, and morphology of mouse epididymal sperm, optimizing the diluent, incubation time, sample concentration, and temperature, using a particle counter (CDA-500) to count and size sperm and a sperm quality analyzer (SQA-IIC) to measure sperm motility, quantified as the sperm motility index (SMI). The optimal conditions consisted of a 30-min incubation in D-MEM (Dulbecco's modified Eagle's medium; considering cost and availability) at 37 degrees C, with 5 x 10(6)cells mL(-1) in the original solution. Furthermore, the influence of formalin fixation, and the correlation between the automated counter and a manual method were investigated. The sample fixation had no marked effect on the sperm count or morphology assessment. A linear correlation was observed between the manual and automated methods (y=0.920x +0.276; r(2)=0.571; p<0.001; range: (3-6) x 10(6)). The suitability of the proposed method was confirmed using spermatozoa prepared from mice treated with the reproductive toxin diethylstilbestrol (DES). Using sperm from the cauda epididymidis on one side per mouse, we confirmed that measurement of these sperm parameters using the two devices was simple, rapid, inexpensive, and reproducible.
Tayama
Kuniaki
K
Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute of Public Health, 3-24-1 Hakunincho, Shinjuku-ku, Tokyo 169-0073, Japan. kuniaki_tayama@member.metro.tokyo.jp
Fujita
Hiroshi
H
Takahashi
Hiroshi
H
Nagasawa
Akemichi
A
Yano
Norio
N
Yuzawa
Katsuhiro
K
Ogata
Akio
A
D000818:Animals; D000831:Animals, Newborn; D004054:Diethylstilbestrol / Q000008:administration & dosage / Q000633:toxicity; D004822:Epididymis / Q000166:cytology / Q000187:drug effects; D005557:Formaldehyde; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010316:Particle Size; D015203:Reproducibility of Results; D012662:Semen Preservation / Q000379:methods; D013076:Sperm Count / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects; D013696:Temperature; D016707:Tissue Fixation
D005557:Formaldehyde; D004054:Diethylstilbestrol