| 1. |
- Evander, Mikael, et al.
(författare)
-
Study of ATP-release from acoustically levitated eryhrocytes
- 2007
-
Ingår i: The proceedings of micro total analysis systems 2007. - 9780979806407 ; 2, s. 1372-1374
-
Konferensbidrag (refereegranskat)abstract
- Erythrocytes (red blood cells) are known to produce large amounts of Adenosine Triphosphate (ATP). It has recently become clear that the ATP-release is part of a mechanism controlling the dilation of the body’s blood vessels. The study of the erythrocyte’s behaviour is complicated by the fact that they respond easily to any physical contact. In this paper we propose a new method for studying the dynamics of the ATP-release by combining non-contact acoustic trapping in a microfluidic chip with a chemiluminiscent assay. Sensitivity levels down to 10 pM were achievable and the ATP-release from a cluster of levitated live erythrocytes was recorded.
|
|
| 2. |
|
|
| 3. |
- Malm, Johan, et al.
(författare)
-
Stardardization Developments for Large Scale Biobanks in Smoking Related Diseases - A Model System for Blood Sample Processing and Storage
- 2013
-
Ingår i: Translational Respiratory Medicine. - : Springer Science and Business Media LLC. - 2213-0802. ; 1
-
Tidskriftsartikel (refereegranskat)abstract
- Biobank samples stored in biobanks give researchers and respiratory healthcare institutions access to datasets of analytes valuable for both diagnostic and research practices. The usefulness of these samples in clinical decision-making is highly dependent on their quality and integrity. New procedures that better preserve sample integrity and reduce degradation are being developed to meet the needs of both present and future biobanking. Hereby we present an automatic sample workflow scheme that is designed to handle high numbers of blood samples. Blood fractions are aliquoted, heat sealed using novel technology, and stored in 384 tube high-density sample arrays. The newly developed 384 biobank rack system is especially suited for preserving identical small aliquots. This technology development allows rapid access to a given sample in the frozen archive while maintaining individual sample integrity with sample tube confinement and quality management. We provide data on robotic processing of clinical samples at -80°C, following initial processing, analysis and shipping between laboratories throughout Europe. Subsequent to unpacking, re-sorting, and storage at these sites, the samples have been returned for analysis. Biomarker analysis of 13 common tests in the clinical chemistry unit of the hospital provides evidence of qualitative and stable logistics using the 384-sample tube system.
|
|
| 4. |
- Rezeli, Melinda, et al.
(författare)
-
Development of an MRM assay panel with application to biobank samples from patients with Myocardial Infarction.
- 2013
-
Ingår i: Journal of Proteomics. - : Elsevier BV. - 1874-3919. ; 87:1, s. 16-25
-
Tidskriftsartikel (refereegranskat)abstract
- As part of a Swedish national cardiological research initiative, the development of a quantitative MRM assay is reported for the quantification of eleven putative cardiovascular disease markers. Within the study, patient samples from the LUNDHEARTGENE biobank were processed and nanoLC-MS/MS analysis were performed together with stable isotope dilution strategy for absolute quantification of the target proteins. Excellent linear regressions were achieved for 9 of the 11 peptides with LOQ ranged in the attomolar range. We have utilized the assay for the screening of plasma samples from patients with chest pain, and performed a comparative analysis of patients with ST Segment Elevation Myocardial Infarction and chest pain due to other causes. The assay demonstrates high reproducibility and correlate with clinical findings. Strong correlations were found for several of the apolipoproteins and their respective lipid subfractions (LDL, HDL or triglycerides). ApoC1, apoC2 and apoE were elevated in patients with STEMI. BIOLOGICAL SIGNIFICANCE: MRM assay were developed for putative cardiovascular disease markers as target proteins, and applied to biobanking sample material. The comparative analysis of patients with ST Segment Elevation Myocardial Infarction and chest pain due to other causes shows elevated levels of apoC1, apoC2 and apoE in patients with STEMI. These observations raise interesting novel hypotheses about the role of apolipoproteins C1, C2 and E in the pathophysiology of acute myocardial infarction, which merits further studies.
|
|
| 5. |
- Bryl-Górecka, Paulina, et al.
(författare)
-
Effect of exercise on the plasma vesicular proteome : A methodological study comparing acoustic trapping and centrifugation
- 2018
-
Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 18:20, s. 3101-3111
-
Tidskriftsartikel (refereegranskat)abstract
- Extracellular vesicles (EVs) are a heterogeneous group of actively released vesicles originating from a wide range of cell types. Characterization of these EVs and their proteomes in the human plasma provides a novel approach in clinical diagnostics, as they reflect physiological and pathological states. However, EV isolation is technically challenging with the current methods having several disadvantages, requiring large sample volumes, and resulting in loss of sample and EV integrity. Here, we use an alternative, non-contact method based on a microscale acoustic standing wave technology. Improved coupling of the acoustic resonator increased the EV recovery from 30% in earlier reports to 80%, also displaying long term stability between experiment days. We report a pilot study, with 20 subjects who underwent physical exercise. Plasma samples were obtained before and 1 h after the workout. Acoustic trapping was compared to a standard high-speed centrifugation protocol, and the method was validated by flow cytometry (FCM). To monitor the device stability, the pooled frozen plasma from volunteers was used as an internal control. A key finding from the FCM analysis was a decrease in CD62E+ (E-selectin) EVs 1 h after exercise that was consistent for both methods. Furthermore, we report the first data that analyse differential EV protein expression before and after physical exercise. Olink-based proteomic analysis showed 54 significantly changed proteins in the EV fraction in response to physical exercise, whereas the EV-free plasma proteome only displayed four differentially regulated proteins, thus underlining an important role of these vesicles in cellular communication, and their potential as plasma derived biomarkers. We conclude that acoustic trapping offers a fast and efficient method comparable with high-speed centrifugation protocols. Further, it has the advantage of using smaller sample volumes (12.5 μL) and rapid contact-free separation with higher yield, and can thus pave the way for future clinical EV-based diagnostics.
|
|
| 6. |
- Evander, Mikael, et al.
(författare)
-
Non-contact acoustic capture of microparticles from small plasma volumes.
- 2015
-
Ingår i: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189 .- 1473-0197. ; 15:12, s. 2588-2596
-
Tidskriftsartikel (refereegranskat)abstract
- Microparticles (MP) are small (100-1000 nm) membrane vesicles shed by cells as a response to activation, stress or apoptosis. Platelet-derived MP (PMP) has been shown to reflect the pathophysiological processes of a range of cardiovascular diseases and there is a potential clinical value in using PMPs as biomarkers, as well as a need to better understand the biology of these vesicles. The current method for isolating MP depends on differential centrifugation steps, which require relatively large sample volumes and have been shown to compromise the integrity and composition of the MP population. We present a novel method for rapid, non-contact capture of PMP in minute sample volumes based on a microscale acoustic standing wave technology. Capture of PMPs from plasma is shown by scanning electron microscopy and flow cytometry. Furthermore, the system is characterized with regards to plasma sample concentration and flow rate. Finally, the technique is compared to a standard differential centrifugation protocol using samples from both healthy controls and ST-elevation myocardial infarction (STEMI) patient samples. The acoustic system is shown to offer a quick and automated setup for extracting microparticles from small sample volumes with higher recovery than a standard differential centrifugation protocol.
|
|
