Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.

Manual microscopic inspection of fixed and stained blood smears has remained the gold standard for Plasmodium parasitemia analysis for over a century. Unfortunately, smear preparation consumes time and reagents, while manual microscopy is skill-dependent and labor-intensive. Here, we demonstrate tha...

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Autores principales:	Paul Lebel, Rebekah Dial, Venkata N P Vemuri, Valentina Garcia, Joseph DeRisi, Rafael Gómez-Sjöberg
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/1a4558849d654dd789cf9c45f1b26387
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spelling	oai:doaj.org-article:1a4558849d654dd789cf9c45f1b263872021-12-02T19:58:07ZLabel-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.1553-734X1553-735810.1371/journal.pcbi.1009257https://doaj.org/article/1a4558849d654dd789cf9c45f1b263872021-08-01T00:00:00Zhttps://doi.org/10.1371/journal.pcbi.1009257https://doaj.org/toc/1553-734Xhttps://doaj.org/toc/1553-7358Manual microscopic inspection of fixed and stained blood smears has remained the gold standard for Plasmodium parasitemia analysis for over a century. Unfortunately, smear preparation consumes time and reagents, while manual microscopy is skill-dependent and labor-intensive. Here, we demonstrate that deep learning enables both life stage classification and accurate parasitemia quantification of ordinary brightfield microscopy images of live, unstained red blood cells. We tested our method using both a standard light microscope equipped with visible and near-ultraviolet (UV) illumination, and a custom-built microscope employing deep-UV illumination. While using deep-UV light achieved an overall four-category classification of Plasmodium falciparum blood stages of greater than 99% and a recall of 89.8% for ring-stage parasites, imaging with near-UV light on a standard microscope resulted in 96.8% overall accuracy and over 90% recall for ring-stage parasites. Both imaging systems were tested extrinsically by parasitemia titration, revealing superior performance over manually-scored Giemsa-stained smears, and a limit of detection below 0.1%. Our results establish that label-free parasitemia analysis of live cells is possible in a biomedical laboratory setting without the need for complex optical instrumentation. We anticipate future extensions of this work could enable label-free clinical diagnostic measurements, one day eliminating the need for conventional blood smear analysis.Paul LebelRebekah DialVenkata N P VemuriValentina GarciaJoseph DeRisiRafael Gómez-SjöbergPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Computational Biology, Vol 17, Iss 8, p e1009257 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Paul Lebel Rebekah Dial Venkata N P Vemuri Valentina Garcia Joseph DeRisi Rafael Gómez-Sjöberg Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
description	Manual microscopic inspection of fixed and stained blood smears has remained the gold standard for Plasmodium parasitemia analysis for over a century. Unfortunately, smear preparation consumes time and reagents, while manual microscopy is skill-dependent and labor-intensive. Here, we demonstrate that deep learning enables both life stage classification and accurate parasitemia quantification of ordinary brightfield microscopy images of live, unstained red blood cells. We tested our method using both a standard light microscope equipped with visible and near-ultraviolet (UV) illumination, and a custom-built microscope employing deep-UV illumination. While using deep-UV light achieved an overall four-category classification of Plasmodium falciparum blood stages of greater than 99% and a recall of 89.8% for ring-stage parasites, imaging with near-UV light on a standard microscope resulted in 96.8% overall accuracy and over 90% recall for ring-stage parasites. Both imaging systems were tested extrinsically by parasitemia titration, revealing superior performance over manually-scored Giemsa-stained smears, and a limit of detection below 0.1%. Our results establish that label-free parasitemia analysis of live cells is possible in a biomedical laboratory setting without the need for complex optical instrumentation. We anticipate future extensions of this work could enable label-free clinical diagnostic measurements, one day eliminating the need for conventional blood smear analysis.
format	article
author	Paul Lebel Rebekah Dial Venkata N P Vemuri Valentina Garcia Joseph DeRisi Rafael Gómez-Sjöberg
author_facet	Paul Lebel Rebekah Dial Venkata N P Vemuri Valentina Garcia Joseph DeRisi Rafael Gómez-Sjöberg
author_sort	Paul Lebel
title	Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
title_short	Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
title_full	Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
title_fullStr	Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
title_full_unstemmed	Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.
title_sort	label-free imaging and classification of live p. falciparum enables high performance parasitemia quantification without fixation or staining.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/1a4558849d654dd789cf9c45f1b26387
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Label-free imaging and classification of live P. falciparum enables high performance parasitemia quantification without fixation or staining.

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