Single Cell Type Specific RNA Isolation and Gene Expression Analysis in Rice Using Laser Capture Microdissection (LCM)-Based Method

Gautam, Vibhav; Chatterjee, Sourav; Sarkar, Ananda K.

doi:10.1007/978-1-0716-1068-8_18

Cited by 5 publications

(3 citation statements)

References 15 publications

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“…FACS, also, generally requires > 10,000 cells as start input, making it unsuitable for rare samples. Laser capture microdissection (LCM) is a technology that accurately separates single cells from tissue samples [20]. After staining tissue sections, they are examined under a microscope for target area selection, and the selected area is isolated using a laser.…”

Section: Single-cell Isolationmentioning

confidence: 99%

Digital PCR for Single-Cell Analysis

Fang,

Liu,

Maiga

et al. 2024

Biosensors

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Single-cell analysis provides an overwhelming strategy for revealing cellular heterogeneity and new perspectives for understanding the biological function and disease mechanism. Moreover, it promotes the basic and clinical research in many fields at a single-cell resolution. A digital polymerase chain reaction (dPCR) is an absolute quantitative analysis technology with high sensitivity and precision for DNA/RNA or protein. With the development of microfluidic technology, digital PCR has been used to achieve absolute quantification of single-cell gene expression and single-cell proteins. For single-cell specific-gene or -protein detection, digital PCR has shown great advantages. So, this review will introduce the significance and process of single-cell analysis, including single-cell isolation, single-cell lysis, and single-cell detection methods, mainly focusing on the microfluidic single-cell digital PCR technology and its biological application at a single-cell level. The challenges and opportunities for the development of single-cell digital PCR are also discussed.

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Section: Single-cell Isolationmentioning

confidence: 99%

Digital PCR for Single-Cell Analysis

Fang,

Liu,

Maiga

et al. 2024

Biosensors

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“…This technology was developed at the National Cancer Institute (NCI), originally for the study of heterogeneous cell populations in tumors [ 38 ]. LCM has been widely used for single-cell analysis [ 39 , 40 , 41 , 42 ], with the advantage of preserving the spatial information of single cells in tissues and facilitating the analysis of cell–cell and cell–environment interactions [ 43 , 44 , 45 ]. However, the main restriction that limits the application of LCM in single-cell sequencing is its low throughput.…”

Section: General Process Of Scrna-seqmentioning

confidence: 99%

Microfluidics Facilitates the Development of Single-Cell RNA Sequencing

Pan

Cao

et al. 2022

Biosensors

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Single-cell RNA sequencing (scRNA-seq) technology provides a powerful tool for understanding complex biosystems at the single-cell and single-molecule level. The past decade has been a golden period for the development of single-cell sequencing, with scRNA-seq undergoing a tremendous leap in sensitivity and throughput. The application of droplet- and microwell-based microfluidics in scRNA-seq has contributed greatly to improving sequencing throughput. This review introduces the history of development and important technical factors of scRNA-seq. We mainly focus on the role of microfluidics in facilitating the development of scRNA-seq technology. To end, we discuss the future directions for scRNA-seq.

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“…Within this cadre of technologies, Laser Capture Microdissection (LCM) [ 4 - 6 ] has proven to be a reliable means of procuring tissue cellular populations, achieving high sensitivity and high precision for multiplex genomic and proteomic profiling [ 4 , 7 - 9 ]. LCM has been used to generate data for a wide variety of research questions, ranging from entomology [ 10 - 12 ], agriculture research [ 13 , 14 ], embryology [ 15 , 16 ], metabolic disease [ 17 - 19 ], heart disease [ 20 , 21 ], neurobiology [ 22 , 23 ], infectious disease [ 24 - 26 ], and cancer [ 6 ] (see below).…”

Section: Introductionmentioning

confidence: 99%

Laser Capture Proteomics: spatial tissue molecular profiling from the bench to personalized medicine

Liotta

Pappalardo

Carpino

et al. 2021

Expert Review of Proteomics

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Introduction: Laser Capture Microdissection (LCM) uses a laser to isolate, or capture, specific cells of interest in a complex heterogeneous tissue section, under direct microscopic visualization. Recently, there has been a surge of publications using LCM for tissue spatial molecular profiling relevant to a wide range of research topics. Areas Covered: We summarize the many advances in tissue Laser Capture Proteomics (LCP) using mass spectrometry for discovery, and protein arrays for signal pathway network mapping. This review emphasizes: a) transition of LCM phosphoproteomics from the lab to the clinic for individualized cancer therapy, and b) the emerging frontier of LCM single cell molecular analysis combining proteomics with genomic, and transcriptomic analysis. The search strategy was based on the combination of MeSH terms with expert refinement. Expert Opinion: LCM is complemented by a rich set of instruments, methodology protocols, and analytical A.I. (artificial intelligence) software for basic and translational research. Resolution is advancing to the tissue single cell level. A vision for the future evolution of LCM is presented. Emerging LCM technology is combining digital and AI guided remote imaging with automation, and telepathology, to a achieve multi-omic profiling that was not previously possible.

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Single Cell Type Specific RNA Isolation and Gene Expression Analysis in Rice Using Laser Capture Microdissection (LCM)-Based Method

Cited by 5 publications

References 15 publications

Digital PCR for Single-Cell Analysis

Digital PCR for Single-Cell Analysis

Microfluidics Facilitates the Development of Single-Cell RNA Sequencing

Laser Capture Proteomics: spatial tissue molecular profiling from the bench to personalized medicine

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