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On chip analysis of CNS lymphoma in cerebrospinal fluid.

Turetsky A, Lee K, Song J, Giedt RJ, Kim E, Kovach AE, Hochberg EP, Castro CM, Lee H, Weissleder R - Theranostics (2015)

Bottom Line: Molecular profiling of central nervous system lymphomas in cerebrospinal fluid (CSF) samples can be challenging due to the paucicellular and limited nature of the samples.The system can detect scant lymphoma cells and quantitate their kappa/lambda immunoglobulin light chain restriction patterns.The approach can be further customized for measurement of additional biomarkers, such as those for differential diagnosis of lymphoma subtypes or for prognosis, as well as for imaging exposure to experimental drugs.

View Article: PubMed Central - PubMed

Affiliation: 1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge St, CPZN 5206, Boston, MA 02114, USA.

ABSTRACT
Molecular profiling of central nervous system lymphomas in cerebrospinal fluid (CSF) samples can be challenging due to the paucicellular and limited nature of the samples. Presented herein is a microfluidic platform for complete CSF lymphoid cell analysis, including single cell capture in sub-nanoliter traps, and molecular and chemotherapeutic response profiling via on-chip imaging, all in less than one hour. The system can detect scant lymphoma cells and quantitate their kappa/lambda immunoglobulin light chain restriction patterns. The approach can be further customized for measurement of additional biomarkers, such as those for differential diagnosis of lymphoma subtypes or for prognosis, as well as for imaging exposure to experimental drugs.

No MeSH data available.


Related in: MedlinePlus

On-Chip Imaging. A 1:1 mixture of DB and Daudi cells were captured and stained on-chip using a cocktail of antibodies: anti-CD19-PE, anti-CD20-PE, anti-Kappa-Brilliant Violet 421, anti-Lambda-Alexa Fluor 647, and anti-Ki-67-Alexa Fluor 488. (A) Low-magnification image shows overall capture site layout and cell heterogeneity. Scale bar: 75 µm. (B) High-resolution images of differential expression of individual markers on the two cell lines. Scale bar: 5 µm.
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Figure 4: On-Chip Imaging. A 1:1 mixture of DB and Daudi cells were captured and stained on-chip using a cocktail of antibodies: anti-CD19-PE, anti-CD20-PE, anti-Kappa-Brilliant Violet 421, anti-Lambda-Alexa Fluor 647, and anti-Ki-67-Alexa Fluor 488. (A) Low-magnification image shows overall capture site layout and cell heterogeneity. Scale bar: 75 µm. (B) High-resolution images of differential expression of individual markers on the two cell lines. Scale bar: 5 µm.

Mentions: We chose to use Daudi and DB cells as a model system for on-chip analysis, since they respectively highly express kappa and lambda light chain. To demonstrate both extracellular and intracellular antigen analysis, we performed on-chip staining of CD19/CD20, kappa/lambda, and Ki-67. We prepared samples by spiking known numbers of DB and Daudi lymphoma cells into artificial CSF (see Methods for details). The cells were then fixed and stained on the chip, and imaged in four channels (Fig. 4; see Supplementary Material: Table S2 for antibody clones and fluorochromes). Fig. 4A shows the overlay of the four imaging channels after a 1:1 mixture of DB and Daudi cells were captured and stained on-chip. Fig. 4B demonstrates high-resolution imaging of individual cells and markers. Although the cell populations appear to be heterogeneous, their restricted kappa/lambda expression can be seen at higher magnification.


On chip analysis of CNS lymphoma in cerebrospinal fluid.

Turetsky A, Lee K, Song J, Giedt RJ, Kim E, Kovach AE, Hochberg EP, Castro CM, Lee H, Weissleder R - Theranostics (2015)

On-Chip Imaging. A 1:1 mixture of DB and Daudi cells were captured and stained on-chip using a cocktail of antibodies: anti-CD19-PE, anti-CD20-PE, anti-Kappa-Brilliant Violet 421, anti-Lambda-Alexa Fluor 647, and anti-Ki-67-Alexa Fluor 488. (A) Low-magnification image shows overall capture site layout and cell heterogeneity. Scale bar: 75 µm. (B) High-resolution images of differential expression of individual markers on the two cell lines. Scale bar: 5 µm.
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Related In: Results  -  Collection

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Figure 4: On-Chip Imaging. A 1:1 mixture of DB and Daudi cells were captured and stained on-chip using a cocktail of antibodies: anti-CD19-PE, anti-CD20-PE, anti-Kappa-Brilliant Violet 421, anti-Lambda-Alexa Fluor 647, and anti-Ki-67-Alexa Fluor 488. (A) Low-magnification image shows overall capture site layout and cell heterogeneity. Scale bar: 75 µm. (B) High-resolution images of differential expression of individual markers on the two cell lines. Scale bar: 5 µm.
Mentions: We chose to use Daudi and DB cells as a model system for on-chip analysis, since they respectively highly express kappa and lambda light chain. To demonstrate both extracellular and intracellular antigen analysis, we performed on-chip staining of CD19/CD20, kappa/lambda, and Ki-67. We prepared samples by spiking known numbers of DB and Daudi lymphoma cells into artificial CSF (see Methods for details). The cells were then fixed and stained on the chip, and imaged in four channels (Fig. 4; see Supplementary Material: Table S2 for antibody clones and fluorochromes). Fig. 4A shows the overlay of the four imaging channels after a 1:1 mixture of DB and Daudi cells were captured and stained on-chip. Fig. 4B demonstrates high-resolution imaging of individual cells and markers. Although the cell populations appear to be heterogeneous, their restricted kappa/lambda expression can be seen at higher magnification.

Bottom Line: Molecular profiling of central nervous system lymphomas in cerebrospinal fluid (CSF) samples can be challenging due to the paucicellular and limited nature of the samples.The system can detect scant lymphoma cells and quantitate their kappa/lambda immunoglobulin light chain restriction patterns.The approach can be further customized for measurement of additional biomarkers, such as those for differential diagnosis of lymphoma subtypes or for prognosis, as well as for imaging exposure to experimental drugs.

View Article: PubMed Central - PubMed

Affiliation: 1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge St, CPZN 5206, Boston, MA 02114, USA.

ABSTRACT
Molecular profiling of central nervous system lymphomas in cerebrospinal fluid (CSF) samples can be challenging due to the paucicellular and limited nature of the samples. Presented herein is a microfluidic platform for complete CSF lymphoid cell analysis, including single cell capture in sub-nanoliter traps, and molecular and chemotherapeutic response profiling via on-chip imaging, all in less than one hour. The system can detect scant lymphoma cells and quantitate their kappa/lambda immunoglobulin light chain restriction patterns. The approach can be further customized for measurement of additional biomarkers, such as those for differential diagnosis of lymphoma subtypes or for prognosis, as well as for imaging exposure to experimental drugs.

No MeSH data available.


Related in: MedlinePlus