A computerized spectrophotometric instrumental system to determine the “vertical velocity” of sperm cells: A novel concept

Abstract

Background:The presently available cell motility‐analyzers measure primarily the “horizontal” velocity and there is no instrument available for “vertical” velocity measurement. This development was based on the turbidimetric method of sperm motility analysis.Methods:Sperm was layered at the bottom of the cuvette containing buffer solution and exposed to the spectrophotometric light path at different heights to track the vertically moving sperms. The vertical movement was materialized with the development of an electromechanical up–down movement devise for the cuvette accomplished with the help of a cuvette holder‐stepper motor‐computer assembly. The entire system was controlled by the necessary motion control, data acquisition, and data processing softwares developed for cuvette movement and data analysis.Results:Using goat sperm as the model a unique computer‐based spectrophotometric system has been developed for the first time to determine the average “vertical” velocity of motile cells.Conclusions:Undertaking upward movement against gravity is much tougher as compared with horizontal movement. Consequently average vertical velocity is expected to be a much better identifying parameter for assessing semen and other motile cell quality. The novel instrumental system developed by us has thus the potential for immense application in human infertility clinics, animal‐breeding centres, centres for conservation of endangered species, and also for research work on vertical velocity of spermatozoa and other motile cells, such as bacteria, protozoa, etc. © 2007 International Society for Analytical Cytology

Purification and Characterization of a Sperm Motility Inhibiting Factor from Caprine Epididymal Plasma

“…Another sophisticated instrument “Computer aided semen analyzer” (CASA) was utlilised that is based on microscopic video photographic method and used for estimating sperm “horizontal” velocity [19] . To determine “vertical” velocity of spermatozoa a unique computer-based spectrophotometric system was developed in our laboratory recently [20] and used for the purpose. This instrument has been named as “SPERMA”.…”

Characterization of a Dipeptidyl Peptidase and its Role in Motility of Rat Epididymal Maturing Spermatozoa

“…The movement of the cells in the vertical plane was measured using the automated computerized spectrophotometric system, 33 as all the motility analyzers till date, including CASA, never considered the importance of motility in the vertical direction. The computerized spectrophotometric assay method showed a drastic inhibition in the percentage of forward motility with time when treated with sitagliptin (1 mM) as compared to the control sperm.…”

“…The forward motility of spermatozoa was quantified by spectrophotometric method, 32,33 which consists of layering 50 µL of freshly extracted cauda epididymal spermatozoa (200 × 10 6 /mL) mixed with 10% Ficoll in a total volume of 0.5 mL RPS medium using a Hamilton syringe at the bottom of a standard cuvette containing 1.3 mL of RPS medium, which was sufficient to cover the entire width of the light beam. Vigorously motile spermatozoa that moved upward in the light beam at any particular time were registered continuously as an increase of absorbance at 545 nm with a Gilford spectrophotometer equipped with a recorder.…”

Ecto-Cyclic AMP Independent Protein Kinase-A Potent Regulator of L-type Calcium Channel and Forward Motility of Goat (Capra indicus) Epididymal Spermatozoa

“…Forward motility of spermatozoa was estimated using spectrophotometric method [41,43,44]. The microscopic method of motility assay described above takes into consideration the number of cells with forward progression but not their velocity, whereas the spectrophotometric method reflects the percentage of forward motile cell as well as their velocity.…”

“…The movement of the cells in the vertical plane was measured using the automated computerized spectrophotometric system [43] as all the motility analyzers till date, including CASA, never considered the importance of motility in vertical direction which is otherwise crucial for fertilization in female system.…”