Morphological Measurement of Living Cells in Methanol with Digital Holographic Microscopy

Abstract

Cell morphology is the research foundation in many applications related to the estimation of cell status, drug response, and toxicity screening. In biomedical field, the quantitative phase detection is an inevitable trend for living cells. In this paper, the morphological change of HeLa cells treated with methanol of different concentrations is detected using digital holographic microscopy. The compact image-plane digital holographic system is designed based on fiber elements. The quantitative phase image of living cells is obtained in combination with numerical analysis. The statistical analysis shows that the area and average optical thickness of HeLa cells treated with 12.5% or 25% methanol reduce significantly, which indicates that the methanol with lower concentration could cause cellular shrinkage. The area of HeLa cells treated with 50% methanol is similar to that of normal cells (P > 0.05), which reveals the fixative effect of methanol with higher concentration. The maximum optical thickness of the cells treated with 12.5%, 25%, and 50% methanol is greater than that of untreated cells, which implies the pyknosis of HeLa cells under the effect of methanol. All of the results demonstrate that digital holographic microscopy has supplied a noninvasive imaging alternative to measure the morphological change of label-free living cells.

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

“…Cell volume changes, resulting from cytotoxic treatments or apoptosis events, have recently been investigated with DH microscopy [11,[21][22][23][24][25]. When cells go in to early apoptosis, the first discernible indication is an increase in the cell phase shift followed by a decrease as the cell eventually dies [21].…”

Quantitative Stain-Free and Continuous Multimodal Monitoring of Wound Healing In Vitro with Digital Holographic Microscopy

“…Recently, it has been demonstrated that DHM provides quantitative monitoring of physiological processes through structural analysis and functional imaging which, for example, gives new insight into signaling of cellular water permeability [19], [20], cell morphology changes due to toxins [21]–[24] and infections [25]–[27] as well as micro-calcification, cancer and inflammation mediated tissue alterations [12], [28] and bacteria and mammalian single cell growth [29]–[32].…”

“…Digital holographic microscopy (DHM), a variant of quantitative phase microscopy, enables not only stain-free quantitative phase contrast imaging but also assessment of cell thickness and tissue density by measuring optical path length delay [16] – [18] . Recently, it has been demonstrated that DHM provides quantitative monitoring of physiological processes through structural analysis and functional imaging which, for example, gives new insight into signaling of cellular water permeability [19] , [20] , cell morphology changes due to toxins [21] – [24] and infections [25] – [27] as well as micro-calcification, cancer and inflammation mediated tissue alterations [12] , [28] and bacteria and mammalian single cell growth [29] – [32] .…”

Super-resolution imaging in digital holography by using dynamic grating with a spatial light modulator

“…DH has growing applications in biological cell imaging, microstructure detection, particle field analysis, and temperature field measurement etc. [1][2][3][4][5][6]. The resolution of a digital holographic system is directly related to the numerical aperture (NA) of the recording device and the wavelength used in the recording.…”