Martinelli U, Daponte P, Abate G.F, Bavaro F, Napolitano M, Mauro F, Grimaldi D, Guglielmelli G,

Rapuano S, Moisa S, Scerbo P, Castello G
 

         

Abstract: This research project aims to produce a tomography system able to acquire 3D images of cells in laminar flow was supported by the Italian Ministry of Education, University and Research (MIUR) under the project FIRB 2001 RBAUOIZMZ5. the hardware innovations regard to (i) the pulses acquisition system, (ii) the optoelectronic single cell triggering system and, (iii) the cells flow chamber. The software innovations regard to (i)  the digital signal processing procedure, (ii) the optimized methodology for classification of the different cell types, (iii) the programmable gain of each channel, and (iv) the image pre-processing to detect the micronuclei into the acquired image of the human lymphocytes.The pre-processing method is pointed out in order to (i) minimize the doubtful detections, and (ii) enhance the confidence that in the rejected images there were not micronuclei. Preliminary exeperimental tests are shown, so to highlight the improved performance of the new measurement device.                                                                                              

     
 

The Acquisition Electronics is able to acquire FC parametric Signals at the same time (90° scatter, 615nm Fluorescence Intensity, Area and Width), simultaneously the electronics processes pulses and converts them in digital format at 4096 levels (12 bits) resolution with a sampling rate of 50 Samples/10 ms/channel.
This means:  

a) the time during which the System is "blind" is T<5ms versus T> 70 ms of most popular flow cytometers;
b) the time resolution of analog pulses is 50 times higher than the typical one; commercial cytometers get 1Sa/10ms

  

The research team designed, developed and realized the prototype of a new generation System which conjugates in one functional Unit an advanced Hi-Sensitivity Flow Cytometer (FC) with  an High Speed, Hi-Res. Optoelectronic device to capture and 3D process live images of flowing BN Cells streamed by a laminar flow. This I-FC system is now ready to begin the final testing and problem fixing phases, as required by the quoted MIUR—FIRB RBAO1ZMZ5 project approved procedures. Finally, a scientific validation and a clinical evaluation report will be realized at National Cancer Institute (UR3) in Naples.

 

The optoelectronic sub-system works on, both Image capturing and the triggered Tachometer module, basing on pulses detected  by the cytometry sub-system. The optoelectronic trigger generator performs the crucial synchronisation with the flowing cells when they were  classified as “event s of interest” at the proper detection point.

Three Software Packages were specifically developed in LabVIEWTM to acquire, to analyse flow cytometric acquired data and to trigger the image acquisition sub-system. These three dedicated software packages were realized basing on an innovative  interdisciplinary approach.