Dynamic quantitative photothermal monitoring of cell death of individual human red blood cells upon glucose depletion

Vasudevan, Srivathsan; Chen, George Chung Kit; Andika, Marta; Agarwal, Shuchi; Chen, Peng; Olivo, Malini

doi:10.1117/1.3484260

Cited by 6 publications

(6 citation statements)

References 28 publications

(34 reference statements)

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“…Stimulators of the channels include osmotic shock and energy depletion (37,59). Ca 2ϩ may activate Ca 2ϩ -sensitive K ϩ channels (10,13) with subsequent exit of KCl together with osmotically obliged water thus leading to cell shrinkage (37).…”

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confidence: 99%

Inhibition of suicidal erythrocyte death by blebbistatin

Lang

Qadri

Zelenak

et al. 2011

American Journal of Physiology-Cell Physiology

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Blebbistatin, a myosin II inhibitor, interferes with myosin-actin interaction and microtubule assembly. By influencing cytoskeletal dynamics blebbistatin counteracts apoptosis of several types of nucleated cells. Even though lacking nuclei and mitochondria, erythrocytes may undergo suicidal cell death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the cell surface. Triggers of eryptosis include energy depletion and osmotic shock, which enhance cytosolic Ca(2+) activity with subsequent Ca(2+)-sensitive cell shrinkage and cell membrane scrambling. The present study explored the effect of blebbistatin on eryptosis. Cell membrane scrambling was estimated from binding of annexin V to phosphatidylserine at the erythrocyte surface, cell volume from forward scatter in fluorescence-activated cell sorting analysis and cytosolic Ca(2+) concentration from Fluo3 fluorescence. Exposure to blebbistatin on its own (1-50 μM) did not significantly modify cytosolic Ca(2+) concentration, forward scatter, or annexin V binding. Glucose depletion (48 h) was followed by a significant increase of Fluo3 fluorescence and annexin V binding, effects significantly blunted by blebbistatin (Fluo3 fluorescence ≥ 25 μM, annexin V binding ≥ 10 μM). Osmotic shock (addition of 550 mM sucrose) again significantly increased Fluo3 fluorescence and annexin binding, effects again significantly blunted by blebbistatin (Fluo3 fluorescence ≥ 25 μM, annexin V binding ≥ 25 μM). The present observations disclose a novel effect of blebbistatin, i.e., an influence on Ca(2+) entry and suicidal erythrocyte death following energy depletion and osmotic shock.

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confidence: 99%

Inhibition of suicidal erythrocyte death by blebbistatin

Lang

Qadri

Zelenak

et al. 2011

American Journal of Physiology-Cell Physiology

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“…Contemplating the experimental result shown in Fig. 13, using average size of the RBC as 6.5 μm [23], the separation between the two peaks is estimated about 2 μm. As the thermal wavefront at 350 ns is only 2 μm, this means only the center portion of the RBC is heated up, but the outer region of the cell is still at room temperature.…”

Section: Discussionmentioning

confidence: 99%

Quantitative photothermal phase imaging of red blood cells using digital holographic photothermal microscope

Vasudevan

Chen²,

Lin

et al. 2015

Appl. Opt.

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Photothermal microscopy (PTM), a noninvasive pump-probe high-resolution microscopy, has been applied as a bioimaging tool in many biomedical studies. PTM utilizes a conventional phase contrast microscope to obtain highly resolved photothermal images. However, phase information cannot be extracted from these photothermal images, as they are not quantitative. Moreover, the problem of halos inherent in conventional phase contrast microscopy needs to be tackled. Hence, a digital holographic photothermal microscopy technique is proposed as a solution to obtain quantitative phase images. The proposed technique is demonstrated by extracting phase values of red blood cells from their photothermal images. These phase values can potentially be used to determine the temperature distribution of the photothermal images, which is an important study in live cell monitoring applications.

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“…On the other hand, for the generation of statistically significant data regarding the incidence of RCD and/or abundancy of RCD specific markers, other approaches that allow quantitative analysis are used. In this regard, RCD is monitored using techniques such as: nuclear magnetic resonance, 3 flow cytometry, 4 imaging approaches, 5,6 photothermy 7 or ultrasound frequency evaluation. 8 Additionally, there are other analytical procedures for performing molecular biology assays that are suitable for cell death evaluation 9 like toxicology tests, 10 viability assays 11 and PCR assays.…”

Section: Introductionmentioning

confidence: 99%