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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

Process design. (A) Summary of overall scheme: paucicellular samples are harvested and captured on the chip without preprocessing. Following on-chip fixation, permeabilization, and immunostaining, the chip is imaged and cytometry is carried out with an in-house image process algorithm. (B) Photograph of lymphocyte capture chip attached to a microslide, showing inlet, debris filter, and capture area, which contains four arrays of 20 × 300 single-cell capture sites.
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Figure 1: Process design. (A) Summary of overall scheme: paucicellular samples are harvested and captured on the chip without preprocessing. Following on-chip fixation, permeabilization, and immunostaining, the chip is imaged and cytometry is carried out with an in-house image process algorithm. (B) Photograph of lymphocyte capture chip attached to a microslide, showing inlet, debris filter, and capture area, which contains four arrays of 20 × 300 single-cell capture sites.

Mentions: Fig. 1A summarizes the procedure for lymphocyte detection and profiling. First, samples are harvested, typically in the range of 1-3 mL. The entire sample is then loaded onto the chip; individual cells are captured in sub-nanoliter traps and on-chip stained for fluorescent imaging. Acquired images are then analyzed with an automatic computational algorithm to generate cell characterization data. The 2 × 4 cm2 chip contains 24,000 staggered, butterfly-shaped traps arranged in four bands of 20 × 300 (Fig. 1B; additional details in Supplementary Material). The capture site architecture was optimized to trap a single lymphocyte, while a 4-µm gap between the butterfly “wings” was incorporated to allow smaller cells, such as erythrocytes, to pass through without being captured (Supplementary Material: Fig. S1). Each trap was self-limiting; once a cell blocks the gap, the structure presents high fluidic resistance, preventing further cell-trapping. To remove cellular debris and aggregates, we also incorporated a column-filter in the sample inlet (Supplementary Material: Fig. S1). The chips were fabricated via standard soft lithography and the estimated cost per chip is <$1. Containing a large number of capturing sites, the chip enables high-throughput analysis. For instance, with typical flow rates of 2-5 mL/hr, target cells could be captured and stained in <1 hour, important for processing clinical samples.


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)

Process design. (A) Summary of overall scheme: paucicellular samples are harvested and captured on the chip without preprocessing. Following on-chip fixation, permeabilization, and immunostaining, the chip is imaged and cytometry is carried out with an in-house image process algorithm. (B) Photograph of lymphocyte capture chip attached to a microslide, showing inlet, debris filter, and capture area, which contains four arrays of 20 × 300 single-cell capture sites.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4440438&req=5

Figure 1: Process design. (A) Summary of overall scheme: paucicellular samples are harvested and captured on the chip without preprocessing. Following on-chip fixation, permeabilization, and immunostaining, the chip is imaged and cytometry is carried out with an in-house image process algorithm. (B) Photograph of lymphocyte capture chip attached to a microslide, showing inlet, debris filter, and capture area, which contains four arrays of 20 × 300 single-cell capture sites.
Mentions: Fig. 1A summarizes the procedure for lymphocyte detection and profiling. First, samples are harvested, typically in the range of 1-3 mL. The entire sample is then loaded onto the chip; individual cells are captured in sub-nanoliter traps and on-chip stained for fluorescent imaging. Acquired images are then analyzed with an automatic computational algorithm to generate cell characterization data. The 2 × 4 cm2 chip contains 24,000 staggered, butterfly-shaped traps arranged in four bands of 20 × 300 (Fig. 1B; additional details in Supplementary Material). The capture site architecture was optimized to trap a single lymphocyte, while a 4-µm gap between the butterfly “wings” was incorporated to allow smaller cells, such as erythrocytes, to pass through without being captured (Supplementary Material: Fig. S1). Each trap was self-limiting; once a cell blocks the gap, the structure presents high fluidic resistance, preventing further cell-trapping. To remove cellular debris and aggregates, we also incorporated a column-filter in the sample inlet (Supplementary Material: Fig. S1). The chips were fabricated via standard soft lithography and the estimated cost per chip is <$1. Containing a large number of capturing sites, the chip enables high-throughput analysis. For instance, with typical flow rates of 2-5 mL/hr, target cells could be captured and stained in <1 hour, important for processing clinical samples.

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