Polychromatic digital holographic microscopy: a quasicoherent-noise-free imaging technique to explore the connectivity of living neuronal networks

Céline Larivière-Loiselle; Erik Bélanger; Pierre P. Marquet

doi:10.1117/1.NPh.7.4.040501

16 October 2020 Polychromatic digital holographic microscopy: a quasicoherent-noise-free imaging technique to explore the connectivity of living neuronal networks

Céline Larivière-Loiselle, Erik Bélanger, Pierre P. Marquet

Author Affiliations +

Neurophotonics, Vol. 7, Issue 4, 040501 (October 2020). https://doi.org/10.1117/1.NPh.7.4.040501

Fig. 1

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(a) Experimental setup injecting the SWLL and AOTF into the DHM using an OF, two mirrors (M1 and M2), and an iris (I). (b) Stack of holograms used to obtain (c) a stack of OPD images which are averaged to obtain (d) the P-DHM image.

Fig. 2

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(a) Single-shot image acquired with 540-nm illumination, (b) control image, and (c) P-DHM image. The white dotted lines in (b) and (c) represent line profiles further analyzed in Fig. 3. (1) and (2) Background section of the control and P-DHM images, respectively. (3) and (4) Neuronal process area of the control and P-DHM images, respectively. (i)–(iii) The white dashed circles highlight cell features and the asterisk identifies a small nerve extension further analyzed in Fig. 4. (d) and (e) Height profiles extracted from the white dotted lines in the bottom left corner of (3) and (4). Scale bar indicates 10 μm.

Fig. 3

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Analysis of the height profile from Figs. 2(b) and 2(c). The mean profiles of (a) the control and (b) P-DHM are shown in black, and the single-shot images used for the frame averaging are displayed in color, corresponding to the wavelength used for acquisition. Black dotted lines correspond to the σb of each specific profile, in this case 5.4 and 1.2 nm for the control case and P-DHM, respectively. The inset in (b) is an enlarged portion of the line profile. (c) σb of the line profile for the (a) control case in green and (b) P-DHM in orange as a function of N. The black line is the 1/N-curve with A chosen as the first data point. (d) σb of another line profile extracted from a different location in the same neuron image, displaying a different behavior as a function of N.

Fig. 4

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