Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy

J. Delpiano; L. Pizarro; C. J. Peddie; M. L. Jones; L. D. Griffin; L. M. Collinson

doi:10.1111/jmi.12700

Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy

Título traducido de la contribución: Detección automatizada de células fluorescentes en secciones de fluorescencia en resina para microscopía electrónica y de luz integrada

J. Delpiano, L. Pizarro, C. J. Peddie, M. L. Jones, L. D. Griffin, L. M. Collinson^*

^*Autor correspondiente de este trabajo

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

9 Citas (Scopus)

Resumen

La tomografía de matriz integrada combina imágenes de fluorescencia y electrón de secciones ultrafinas en un microscopio y permite una correlación precisa de alta resolución de proteínas fluorescentes con orgánulos y membranas celulares. Se pueden obtener imágenes de un gran número de secciones en serie de forma secuencial para producir volúmenes alineados a partir de ambas modalidades de imagen, lo que produce enormes cantidades de datos que deben manipularse y procesarse mediante técnicas novedosas. Aquí, presentamos un esquema para la detección automatizada de células fluorescentes dentro de secciones delgadas de resina, que luego podría usarse para impulsar la adquisición automatizada de imágenes de electrones de las regiones objetivo a través de "seguimiento inteligente". El objetivo de este trabajo es ayudar en la optimización del proceso de adquisición de datos a través de la automatización, liberando al operador para trabajar en otras tareas y acelerando el proceso, mientras se reducen las tasas de datos adquiriendo solo imágenes de las regiones de interés. Se ha demostrado que este nuevo método es robusto contra el ruido y capaz de hacer frente a regiones de baja fluorescencia.

Título traducido de la contribución	Detección automatizada de células fluorescentes en secciones de fluorescencia en resina para microscopía electrónica y de luz integrada
Idioma original	Inglés
Páginas (desde-hasta)	109-119
Número de páginas	11
Publicación	Journal of Microscopy
Volumen	271
N.º	1
DOI	https://doi.org/10.1111/jmi.12700
Estado	Publicada - jul. 2018

Nota bibliográfica

Publisher Copyright:
© 2018 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.

Palabras clave

Segmentación celular
Microscopía correlativa de luz y electrónica
Procesamiento de imágenes
Fluorescencia en resina
Microscopía electrónica y óptica integrada
Algoritmo de cuenca

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10.1111/jmi.12700

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title = "Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy",

abstract = "Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high-resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via {\textquoteleft}smart tracking{\textquoteright}. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence.",

keywords = "Cell segmentation, correlative light and electron microscopy, image processing, in-resin fluorescence, integrated light and electron microscopy, watershed algorithm, Segmentaci{\'o}n celular, Microscop{\'i}a correlativa de luz y electr{\'o}nica, Procesamiento de im{\'a}genes, Fluorescencia en resina, Microscop{\'i}a electr{\'o}nica y {\'o}ptica integrada, Algoritmo de cuenca",

author = "J. Delpiano and L. Pizarro and Peddie, {C. J.} and Jones, {M. L.} and Griffin, {L. D.} and Collinson, {L. M.}",

note = "Funding Information: This work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001999), the UK Medical Research Council (FC001999) and the Wellcome Trust (FC001999); by the MRC, BBSRC and EPSRC under grant award MR/K01580X/1 to LMC; and by Fondo de Ayuda a la Investigacion (FAI), Universidad de los Andes, under grant INV-IN-2017-05 to J.D. Publisher Copyright: {\textcopyright} 2018 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.",

year = "2018",

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AU - Griffin, L. D.

AU - Collinson, L. M.

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N2 - Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high-resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via ‘smart tracking’. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence.

AB - Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high-resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via ‘smart tracking’. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence.

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Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy

Resumen

Nota bibliográfica

Palabras clave

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