On-chip cell imaging flow cytometry by high-speed multicolor optoacoustic microscopy (Conference Presentation)

2020 ◽  
Author(s):  
Lei Xi
Keyword(s):  
Flow Cytometry ◽  
High Speed ◽  
Cell Imaging ◽  
Imaging Flow Cytometry ◽  
On Chip

scholarly journals Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering

2019 ◽  
Vol 116 (32) ◽  
pp. 15842-15848 ◽  
Author(s):  
Yuta Suzuki ◽  
Koya Kobayashi ◽  
Yoshifumi Wakisaka ◽  
Dinghuan Deng ◽  
Shunji Tanaka ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Raman Scattering ◽  
Cancer Biology ◽  
High Speed ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Imaging Flow Cytometry ◽  
Stimulated Raman

Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.

scholarly journals On-chip light-sheet fluorescence imaging flow cytometry at a high flow speed of 1 m/s

2018 ◽  
Vol 9 (7) ◽  
pp. 3424 ◽  
Author(s):  
Taichi Miura ◽  
Hideharu Mikami ◽  
Akihiro Isozaki ◽  
Takuro Ito ◽  
Yasuyuki Ozeki ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Fluorescence Imaging ◽  
Light Sheet ◽  
Flow Speed ◽  
High Flow ◽  
Imaging Flow Cytometry ◽  
On Chip

Optofluidic time-stretch imaging – an emerging tool for high-throughput imaging flow cytometry

Lab on a Chip ◽  
2016 ◽  
Vol 16 (10) ◽  
pp. 1743-1756 ◽  
Author(s):  
Andy K. S. Lau ◽  
Ho Cheung Shum ◽  
Kenneth K. Y. Wong ◽  
Kevin K. Tsia
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Cell Imaging ◽  
Imaging Flow Cytometry ◽  
Single Cell Imaging ◽  
Optical Time ◽  
High Throughput Imaging

Optical time-stretch imaging is now proven for ultrahigh-throughput optofluidic single-cell imaging, at least 10–100 times faster.

RhoA Gtpase Regulates Erythroblast Cytokinesis and Enucleation By Dynamic Coordination Of The Microtubule-Actomyosin Machineries For Successful Abscission

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 311-311
Author(s):  
Diamantis G. Konstantinidis ◽  
Suvarnamala Pushkaran ◽  
Katie M. Giger ◽  
Ping Zhou ◽  
Paul R. Andreassen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Light Chain ◽  
High Speed ◽  
Cell Imaging ◽  
Vesicular Transport ◽  
Annexin V ◽  
Regulatory Light Chain ◽  
Myosin Regulatory Light Chain ◽  
Dynamic Coordination ◽  
Rhoa Gtpase

Abstract RhoA GTPase is known to regulate cell adhesion, actomyosin cytoskeleton, and cytokinesis. Specifically, it contributes to cytokinesis in sea urchin embryos, Xenopus and HeLa cells by associating with the microtubule ends at the abscission area marking the position for the daughter cell separation and by activating myosin through phosphorylation of the myosin regulatory light chain (MRLC) (Piekny et al, Trends in Cell Biology 2005). Erythroblast enucleation, the process through which orthochromatic erythroblasts expel their nucleus during the final stage of mammalian erythropoiesis, is a multi-step process resembling asymmetric cytokinesis (reviewed by Li, Developmental Cell 2013). It requires establishment of cell polarity through microtubule function, followed by formation of a contractile actomyosin ring between reticulocyte and pyrenocyte (Konstantinidis et al, Blood 2012). To define the mechanistic contribution of RhoA signaling in terminal erythroid maturation, we generated mice with erythroid specific RhoA deletion (EpoR-GFPcre-driven). Erythroid-specific deletion of RhoA, confirmed in the CD71-positive cells by PCR and immunoblotting, caused severe anemia, fatal in utero by E15.5. RhoA-deficient peripheral primitive blood cells were large with significant poikilocytosis and anisocytosis, frequently binuclear or multinuclear with polyploidy of the genetic material as demonstrated by flow cytometry with propidium iodide, and with incomplete clearance of the Golgi network and mitochondria. RhoAΔ/Δ fetal livers had progressively decreased cellularity. The erythroblast populations as determined by CD71-Ter119-FSC flow cytometry as well as by multispectral high-speed cell imaging in flow, demonstrated progressive decline and significantly decreased reticulocyte production (n=3, p<0.05), compatible with a fatal intrauterine anemia. RhoAΔ/Δ definitive erythroblasts also exhibited increased frequency of polyploidy. These polyploid cells were Ki67-negative by immunohistochemistry, indicating that they were not actively mitotic. Apoptosis was not significantly increased in the RhoA-deficient erythroblasts as determined by Annexin-V positivity. However, the late RhoAΔ/Δ erythroblasts demonstrated a significant increase in necrosis as determined by 7AAD and Annexin-V double-positivity (n=3, p<0.05). Increased phosphorylation of p53 was evident by immunoblotting, likely functioning as a polyploidy checkpoint after failure of abscission due to RhoA deficiency. Further evidence that RhoA is significant for microtubule organization in erythroblasts was attained by immuno-localization of RhoA at the microtubule ends and by decreased nuclear polarization in EpoR-Cre RhoAΔ/Δ orthochromatic erythroblasts, as shown by decreased delta centroid Ter119-Draq5 (the distance between the centers of the erythroblast and the nucleus) in multispectral high-speed cell imaging in flow. Phosphorylated myosin regulatory light chain (pMRLC) was found significantly decreased by immunoblotting of RhoAΔ/Δ fetal liver erythroblasts, indicating that RhoA controls phosphorylation of MRLC in erythropoiesis. Increased PAK phosphorylation was noted pointing towards compensatory upregulation of Rac GTPases in the RhoA-deficient cells, which was confirmed by quantitative RT-PCR. We further examined the erythroblastic islands in RhoAΔ/Δfetal liver by transmission electron microscopy and found that RhoA-deficient erythroblasts, frequently dysplastic and binucleated, were loosely associated with the central macrophage and there was a paucity of intracellular vacuoles in comparison to WT erythroblasts, indicating possible defects in vesicular transport, which has also been shown to play a role in erythroblast enucleation (Keerthivasan et al, Blood 2010). Thus, RhoA GTPase regulates erythroblast differentiation and enucleation by dynamic coordination of the microtubule-actomyosin machineries to achieve successful abscission. In addition, RhoA may also regulate erythroblast adhesive interactions with the central macrophage in erythroblastic islands and intracellular vesicular transport. Our results reveal novel molecular components and cellular processes in erythropoiesis that may be important for improving the efficiency of red blood cell production in vitro. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Sickle Cell Imaging Flow Cytometry Assay (SIFCA)

2015 ◽  
pp. 279-292 ◽  
Author(s):  
Kleber Y. Fertrin ◽  
Leigh Samsel ◽  
Eduard J. van Beers ◽  
Laurel Mendelsohn ◽  
Gregory J. Kato ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Sickle Cell ◽  
Cell Imaging ◽  
Flow Cytometry Assay ◽  
Imaging Flow Cytometry

An optofluidic system with integrated microlens arrays for parallel imaging flow cytometry

Lab on a Chip ◽  
2018 ◽  
Vol 18 (23) ◽  
pp. 3631-3637 ◽  
Author(s):  
Gregor Holzner ◽  
Ying Du ◽  
Xiaobao Cao ◽  
Jaebum Choo ◽  
Andrew J. deMello ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Speed ◽  
Parallel Imaging ◽  
Single Cells ◽  
Rapid Analysis ◽  
High Speed Imaging ◽  
Microlens Arrays ◽  
Imaging Flow Cytometry

In recent years, high-speed imaging has become increasingly effective for the rapid analysis of single cells in flowing environments.

scholarly journals Development of On-Chip Multi-Imaging Flow Cytometry for Identification of Imaging Biomarkers of Clustered Circulating Tumor Cells

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e104372 ◽  
Author(s):  
Hyonchol Kim ◽  
Hideyuki Terazono ◽  
Yoshiyasu Nakamura ◽  
Kazuko Sakai ◽  
Akihiro Hattori ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Imaging Biomarkers ◽  
Imaging Flow Cytometry ◽  
On Chip

Deep learning‐enabled imaging flow cytometry for high‐speed Cryptosporidium and Giardia detection

2021 ◽  
Author(s):  
Shaobo Luo ◽  
Kim Truc Nguyen ◽  
Binh T. T. Nguyen ◽  
Shilun Feng ◽  
Yuzhi Shi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Deep Learning ◽  
High Speed ◽  
Imaging Flow Cytometry

scholarly journals Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip

2011 ◽  
Vol 12 (6) ◽  
pp. 3618-3634 ◽  
Author(s):  
Masahito Hayashi ◽  
Akihiro Hattori ◽  
Hyonchol Kim ◽  
Hideyuki Terazono ◽  
Tomoyuki Kaneko ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Imaging Flow Cytometry ◽  
On Chip
Sign in / Sign up

Export Citation Format

Share Document