Use of laser microdissection greatly improves the recovery of DNA from sperm on microscope slides

Elliott, Keith; Hill, D.S; Lambert, Cathy; Burroughes, T.R; Gill, Peter

doi:10.1016/s0379-0738(03)00267-6

Cited by 120 publications

(39 citation statements)

References 15 publications

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“…Compared to other alternative methods, 4,7,8 the proposed microdevice has the following four advantages: (1) because this microfluidic chip does not contain any extra components such as microelectrodes or piezo transducers, the fabrication process is simple and the cost is low; (2) the device eliminates the need for complicated external instruments, aside from two pumps for fluid control, making the device easy to automate and miniaturize; (3) the chip material (PDMS) can be replaced with polymethyl methacrylate or other polymer materials that are easy for mass production, 28 and thus the microfluidic chip has a great potential to be transformed into a disposable product for forensic analysis of sexual assault cases; and (4) the microdevice is easily integrated, and the integration of this microfluidic chip with subsequent sample processing chips (e.g., DNA extraction, PCR, or STR chips 29,30 ) may offer a chance to replace the conventional labor-consuming and time-consuming processes for forensic evidence analysis.…”

Section: Separation Performance Analysismentioning

confidence: 99%

“…Techniques such as filtration 6 and dielectrophoresis (DEP) 7 have also been used for separation, but the purity (<80% male fraction) of the product is low. Other techniques, such as laser microdissection 8,9 and antibody-based fluorescence-activated cell sorting (FACS), 10 have been reported as well. These methods are suitable for samples with low sperm concentrations, but their costs are relatively high.…”

Section: Introductionmentioning

confidence: 99%

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Separation of sperm and epithelial cells based on the hydrodynamic effect for forensic analysis

et al. 2015

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In sexual assault cases, forensic samples are a mixture of sperm from the perpetrator and epithelial cells from the victim. To obtain an independent short tandem repeat (STR) profile of the perpetrator, sperm cells must be separated from the mixture of cells. However, the current method used in crime laboratories, namely, differential extraction, is a time-consuming and labor-intensive process. To achieve a rapid and automated sample pretreatment process, we fabricated a microdevice for hydrodynamic and size-based separation of sperm and epithelial cells. When cells in suspension were introduced into the device's microfluidic channels, they were forced to flow along different streamlines and into different outlets due to their different diameters. With the proposed microdevice, sperm can be separated within a short period of time (0.5 h for a 50-ll mock sample). The STR profiles of the products in the sperm outlet reservoir demonstrated that a highly purified male DNA fraction could be obtained (94.0% male fraction). This microdevice is of low-cost and can be easily integrated with other subsequent analysis units, providing great potential in the process of analyzing sexual assault evidence as well as in other areas requiring cell sorting. V C 2015 AIP Publishing LLC.

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Section: Separation Performance Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Separation of sperm and epithelial cells based on the hydrodynamic effect for forensic analysis

et al. 2015

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“…Since primary pancreatic cancer is characterized by its strong desmoplasia and inflammatory reactions, it is difficult to obtain near-pure cancer cells. Laser microdissection has been demonstrated to be a powerful tool for isolating specific target cells from more complex tissues and from smears in which different kinds of cells are present [9]. …”

Section: Introductionmentioning

confidence: 99%

Integrated Analysis of Copy Number Alterations and Loss of Heterozygosity in Human Pancreatic Cancer Using a High-Resolution, Single Nucleotide Polymorphism Array

Lin

Asaoka²,

Tada³

et al. 2008

Oncology

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Objective: To chart molecular genetic events in pancreatic cancer. Methods: We analyzed genome-wide copy number alterations and loss of heterozygosity (LOH) in 25 established pancreatic cancer cell lines using a high-density single nucleotide polymorphism (SNP) array. We verified the data using genomic PCR and applied them to clinical samples. Results: Twenty-six homozygous deletion regions were detected in at least 1 cell line and LOH was found at 9p, 18q, 17p, 8p, 13q, 6q, 3p, 6p, 22q, 9q and 12q with high frequency (>50%), consistent with a previous study. Moreover, we found 23 amplified regions in at least 2 cell lines, including 8 unreported loci. We then examined representative genes at the 8 amplified loci in matched pairs of pancreatic cancer and normal tissues. The amplification was detected in 1 (7.1%) to 5 (35.7%) of 14 microdissected tissue specimens. Conclusion: Using high-resolution SNP arrays, we studied genome-wide copy number alterations and LOH simultaneously. We identified several novel and minute genomic amplifications, which contained candidate oncogenes in human pancreatic cancers.

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“…DNA analysis has become the prime tool for identification of the source of biological traces [54]. Most reports focus on the use of LAM to isolate a single sperm from a vaginal smear for genetic analysis in the investigation of sexual assault, the first application described by Elliott et al [55]. An even more challenging situation occurs when cells of a victim are much more abundant than the cells of the perpetrator.…”

Section: Laser-assisted Microdissectionmentioning

confidence: 99%

The Laser Technology: New Trends in Biology and Medicine

Legrès¹,

Chamot²,

Varna³

et al. 2014

JMP

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In fifty years, laser technology has made great progress, and its many applications make it essential in everyday life. However, this technology is still open to numerous developments. Across multiple applications, there is particular focus in the field of medicine, for diagnosis for tailored therapies, and as a research tool in biology. Whereas its use is now well-demonstrated in ophthalmologic and dermatologic treatments, and surgery, one of the most fascinating aspects of laser technology in the field of biology emerged in the late 1990s with the development of devices able to perform fine dissections of biological tissues using a laser beam. The so-called laser-associated microdissection offers a rapid, precise method of isolating and removing targeted cells or groups of cells from complex biological tissues. It represents the missing link between clinical observations and the intrinsic physiological mechanisms of biological tissues. The molecular examination of pathologically altered cells and tissues for DNA, RNA, and protein expression has revolutionized research and diagnosis in pathology, enabling assessment of the role of the cell type in the normal physiological or disease process. Alongside conventional diagnostic and therapeutic approaches, another field of application contribute to the development of targeted treatments at the nanoscale level of laser technology, mainly in the field of cancer, leading to design new and innovative strategies in drug delivery and image-guided surgery. Most of these approaches, but although not exhaustively, will be presented here.

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Use of laser microdissection greatly improves the recovery of DNA from sperm on microscope slides

Cited by 120 publications

References 15 publications

Separation of sperm and epithelial cells based on the hydrodynamic effect for forensic analysis

Separation of sperm and epithelial cells based on the hydrodynamic effect for forensic analysis

Integrated Analysis of Copy Number Alterations and Loss of Heterozygosity in Human Pancreatic Cancer Using a High-Resolution, Single Nucleotide Polymorphism Array

The Laser Technology: New Trends in Biology and Medicine

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