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Non‐invasive, label‐free cell counting and quantitative analysis of adherent cells using digital holography
Author(s) -
MÖLDER A.,
SEBESTA M.,
GUSTAFSSON M.,
GISSELSON L.,
WINGREN A. GJÖRLOFF,
ALM K.
Publication year - 2008
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2008.02095.x
Subject(s) - hemocytometer , holography , digital holographic microscopy , cell counting , phase imaging , digital holography , microscope , computer science , microscopy , optics , repeatability , phase (matter) , optical microscope , biomedical engineering , artificial intelligence , computer vision , materials science , cell , mathematics , chemistry , physics , pathology , statistics , medicine , scanning electron microscope , biochemistry , cell cycle , organic chemistry
Summary Manual cell counting is time consuming and requires a high degree of skill on behalf of the person performing the count. Here we use a technique that utilizes digital holography, allowing label‐free and completely non‐invasive cell counting directly in cell culture vessels with adherent viable cells. The images produced can provide both quantitative and qualitative phase information from a single hologram. The recently constructed microscope Holomonitor™ (Phase Holographic Imaging AB, Lund, Sweden) combines the commonly used phase contrast microscope with digital holography, the latter giving us the possibility of achieving quantitative information on cellular shape, area, confluence and optical thickness. This project aimed at determining the accuracy and repeatability of cell counting measurements using digital holography compared to the conventional manual cell counting method using a haemocytometer. The collected data were also used to determine cell size and cellular optical thickness. The results show that digital holography can be used for non‐invasive automatic cell counting as precisely as conventional manual cell counting