LEGO bricks used as chemotactic chambers: evaluation by a computer-assisted image analysis technique.
One of the main techniques used to explore neutrophil motility, employs micropore filters in chemotactic chambers. Many new models have been proposed, in order to perform multiple microassays in a rapid, inexpensive and reproducible way. In this work, LEGO bricks have been used as chemotactic chambers in the evaluation of neutrophil random motility and chemotaxis and compared with conventional Boyden chambers in a "time-response" experiment. Neutrophil motility throughout the filters was evaluated by means of an image-processing workstation, in which a dedicated algorithm recognizes and counts the cells in several fields and focal planes throughout the whole filter; correlates counts and depth values; performs a statistical analysis of data; calculates the true value of neutrophil migration; determines the distribution of cells; and displays the migration pattern. By this method, we found that the distances travelled by the cells in conventional chambers and in LEGO bricks were perfectly identical, both in random migration and under chemotactic conditions. Moreover, no interference with the physiological behaviour of neutrophils was detectable. In fact, the kinetics of migration was identical both in random migration (characterized by a gaussian pattern) and in chemotaxis (characterized by a typical stimulation peak, previously identified by our workstation). In conclusion, LEGO bricks are extremely precise devices. They are simple to use and allow the use of small amounts of chemoattractant solution and cell suspension, supplying by itself a triplicate test. LEGO bricks are inexpensive, fast and suitable for current diagnostic activity or for research investigations in every laboratory.
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Unit of Haematology, Department of Oncology, Transplants and Advanced Technologies in Medicine, University of Pisa, Pisa, Italy. a.azzara@do.med.unipi.it
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D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D002633:Chemotaxis / Q000502:physiology*; D015194:Diffusion Chambers, Culture / Q000295:instrumentation*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008851:Micropore Filters; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*