High-throughput, label-free and slide-free histological imaging by computational microscopy and unsupervised learning

Rapid and high-resolution histological imaging with minimal tissue preparation has long been a challenging and yet captivating medical pursue. Here, we propose a promising and transformative histological imaging method, termed computational high-throughput autofluorescence microscopy by pattern illumination (CHAMP). With the assistance of computational microscopy, CHAMP enables high-throughput and label-free imaging of thick and unprocessed tissues with large surface irregularity at an acquisition speed of 10 mm2/10 seconds with 1.1-um lateral resolution. Moreover, the CHAMP image can be transformed into a virtually stained histological image (Deep-CHAMP) through unsupervised learning within 15 seconds, where significant cellular features are quantitatively extracted with high accuracy. The versatility of CHAMP is experimentally demonstrated using mouse brain/kidney tissues prepared with various clinical protocols, which enables a rapid and accurate intraoperative/postoperative pathological examination without tissue processing or staining, demonstrating its great potential as an assistive imaging platform for surgeons and pathologists to provide optimal adjuvant treatment.

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High‐Throughput, Label‐Free and Slide‐Free Histological Imaging by Computational Microscopy and Unsupervised Learning

Advanced Science ◽

10.1002/advs.202102358 ◽

2021 ◽

pp. 2102358

Author(s):

Yan Zhang ◽

Lei Kang ◽

Ivy H. M. Wong ◽

Weixing Dai ◽

Xiufeng Li ◽

...

Keyword(s):

Unsupervised Learning ◽

High Throughput ◽

Label Free

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High‐Throughput, Label‐Free and Slide‐Free Histological Imaging by Computational Microscopy and Unsupervised Learning (Adv. Sci. 2/2022)

Advanced Science ◽

10.1002/advs.202270012 ◽

2022 ◽

Vol 9 (2) ◽

pp. 2270012

Author(s):

Yan Zhang ◽

Lei Kang ◽

Ivy H. M. Wong ◽

Weixing Dai ◽

Xiufeng Li ◽

...

Keyword(s):

Unsupervised Learning ◽

High Throughput ◽

Label Free

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Real-time imaging of cellular forces using optical interference

Nature Communications ◽

10.1038/s41467-021-23734-4 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Andrew T. Meek ◽

Nils M. Kronenberg ◽

Andrew Morton ◽

Philipp Liehm ◽

Jan Murawski ◽

...

Keyword(s):

Cell Culture ◽

Real Time ◽

High Throughput ◽

Mechanical Activity ◽

Dynamic Processes ◽

Imaging Method ◽

Neonatal Cardiomyocytes ◽

A Cell ◽

Cellular Forces ◽

Time Acquisition

AbstractImportant dynamic processes in mechanobiology remain elusive due to a lack of tools to image the small cellular forces at play with sufficient speed and throughput. Here, we introduce a fast, interference-based force imaging method that uses the illumination of an elastic deformable microcavity with two rapidly alternating wavelengths to map forces. We show real-time acquisition and processing of data, obtain images of mechanical activity while scanning across a cell culture, and investigate sub-second fluctuations of the piconewton forces exerted by macrophage podosomes. We also demonstrate force imaging of beating neonatal cardiomyocytes at 100 fps which reveals mechanical aspects of spontaneous oscillatory contraction waves in between the main contraction cycles. These examples illustrate the wider potential of our technique for monitoring cellular forces with high throughput and excellent temporal resolution.

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Development of a Robust High-Throughput Screening Platform for Inhibitors of the Striatal-Enriched Tyrosine Phosphatase (STEP)

International Journal of Molecular Sciences ◽

10.3390/ijms22094417 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4417

Author(s):

Lester J Lambert ◽

Stefan Grotegut ◽

Maria Celeridad ◽

Palak Gosalia ◽

Laurent JS De Backer ◽

...

Keyword(s):

Drug Discovery ◽

High Throughput ◽

Tyrosine Kinases ◽

High Throughput Screening ◽

Kinase Inhibitors ◽

Tyrosine Phosphatase ◽

Protein Tyrosine Phosphatases ◽

Synaptic Function ◽

Label Free ◽

Protein Tyrosine

Many human diseases are the result of abnormal expression or activation of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Not surprisingly, more than 30 tyrosine kinase inhibitors (TKIs) are currently in clinical use and provide unique treatment options for many patients. PTPs on the other hand have long been regarded as “undruggable” and only recently have gained increased attention in drug discovery. Striatal-enriched tyrosine phosphatase (STEP) is a neuron-specific PTP that is overactive in Alzheimer’s disease (AD) and other neurodegenerative and neuropsychiatric disorders, including Parkinson’s disease, schizophrenia, and fragile X syndrome. An emergent model suggests that the increase in STEP activity interferes with synaptic function and contributes to the characteristic cognitive and behavioral deficits present in these diseases. Prior efforts to generate STEP inhibitors with properties that warrant clinical development have largely failed. To identify novel STEP inhibitor scaffolds, we developed a biophysical, label-free high-throughput screening (HTS) platform based on the protein thermal shift (PTS) technology. In contrast to conventional HTS using STEP enzymatic assays, we found the PTS platform highly robust and capable of identifying true hits with confirmed STEP inhibitory activity and selectivity. This new platform promises to greatly advance STEP drug discovery and should be applicable to other PTP targets.

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High-Throughput Label-Free Biochemical Assays Using Infrared Matrix-Assisted Desorption Electrospray Ionization Mass Spectrometry

Analytical Chemistry ◽

10.1021/acs.analchem.1c00737 ◽

2021 ◽

Vol 93 (17) ◽

pp. 6792-6800

Author(s):

Fan Pu ◽

Andrew J. Radosevich ◽

James W. Sawicki ◽

David Chang-Yen ◽

Nari N. Talaty ◽

...

Keyword(s):

Mass Spectrometry ◽

Electrospray Ionization ◽

High Throughput ◽

Electrospray Ionization Mass Spectrometry ◽

Desorption Electrospray Ionization ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Label Free ◽

Biochemical Assays

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Development of a Novel Label-Free and High-Throughput Arginase-1 Assay Using Self-Assembled Monolayer Desorption Ionization Mass Spectrometry

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/24725552211000677 ◽

2021 ◽

pp. 247255522110006

Author(s):

Michael D. Scholle ◽

Zachary A. Gurard-Levin

Keyword(s):

Mass Spectrometry ◽

Drug Discovery ◽

High Throughput ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Label Free ◽

Desorption Ionization ◽

Arginase 1 ◽

Self Assembled ◽

High Throughput Assay

Arginase-1, an enzyme that catalyzes the reaction of L-arginine to L-ornithine, is implicated in the tumor immune response and represents an interesting therapeutic target in immuno-oncology. Initiating arginase drug discovery efforts remains a challenge due to a lack of suitable high-throughput assay methodologies. This report describes the combination of self-assembled monolayers and matrix-assisted laser desorption ionization mass spectrometry to enable the first label-free and high-throughput assay for arginase activity. The assay was optimized for kinetically balanced conditions and miniaturized, while achieving a robust assay (Z-factor > 0.8) and a significant assay window [signal-to-background ratio > 20] relative to fluorescent approaches. To validate the assay, the inhibition of the reference compound nor-NOHA (Nω-hydroxy-nor-L-arginine) was evaluated, and the IC50 measured to be in line with reported results (IC50 = 180 nM). The assay was then used to complete a screen of 175,000 compounds, demonstrating the high-throughput capacity of the approach. The label-free format also eliminates opportunities for false-positive results due to interference from library compounds and optical readouts. The assay methodology described here enables new opportunities for drug discovery for arginase and, due to the assay flexibility, can be more broadly applicable for measuring other amino acid–metabolizing enzymes.

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