The Design And Development Of The Built-in Cell Dynamic 3D Microscopic Imaging System In The Incubator

The rapid development of cell biology has put forward new requirements for the microscopic observation methods of cells.Nowadays,it has been a key experimental technique in biological basic research and related applied research to observe and record live cell growth processes in real time and in long term.However,microscopy instruments with these capabilities are nowhere to find.The technical bottleneck lies in that the traditional optical microscope adopts fixed Kohler illumination for bright field imaging,resulting that it is impossible to observe the unstained cells directly.Moreover,it cannot provide essential environmental conditions for live cells growth,resulting that it is difficult to observe the cells in an active state.In response to the above problems,we have developed a dynamic monitoring system for three-dimensional information of live cells in incubator and named it Cellmonitor.It consists of a microscope and supporting operating software.It employs a programmable LED array with adjustable illumination aperture and angle as the micro illumination source,which enables Cellmonitor to realize multi-mode imaging functions such as bright field,dark field and Rheinberg imaging under the same system.By using self-developed differential phase contrast quantitative phase imaging algorithm in optimal illumination mode,with the aid of LED arrays for coded illumination of the sample,Cellmonitor realizes real-time quantitative threedimensional imaging of samples such as unstained cells under non-interference conditions.The device uses an original miniaturized microscopic imaging system design,which enables it to be placed directly into the incubator for long-term,stable observation of the morphological development of cells.The software integrates many image analysis functions such as cell segmentation counting,which enables Cellmonitor to realize multi-dimensional image acquisition and growth parameter analysis of live cells efficiently.This paper completed imaging algorithm design,optical design,mechanical structure design,hardware driver development,and operation software development to form a complete set of equipment with high completion.Finally,we tested the functions and performance of the system by using samples such as Hela cells to verify its reliability.