ABSTRACT/SYNOPSIS The recent identification of rare cell populations within tissues that are associated with specific biological behaviors, e.g., progenitor cells, has illuminated a limitation of current technologies to study such adherent cells directly from primary tissues. The micropallet array is a recently developed technology designed to address this limitation by virtue of its capacity to isolate and recover single adherent cells on individual micropallets. The capacity to apply this technology to primary tissues and cells with restricted growth characteristics, particularly adhesion requirements, is critically dependent upon the capacity to generate functional extracellular matrix (ECM) coatings. The discontinuous nature of the micropallet array surface provides specific constraints on the processes for generating the desired ECM coatings that are necessary to achieve the full functional capacity of the micropallet array. We have developed strategies, reported herein, to generate functional coatings with various ECM protein components: fibronectin, EHS tumor basement membrane extract, collagen, and laminin-5; confirmed by evaluation for rapid cellular adherence of four dissimilar cell types: fibroblast, breast epithelial, pancreatic epithelial, and myeloma. These findings are important for the dissemination and expanded use of micropallet arrays and similar microtechnologies requiring the integrated use of ECM protein coatings to promote cellular adherence.
We present a magnetic micropallet array and demonstration of its capacity to recover specific, individual adherent cells from large populations and deliver them for downstream single cell analysis. A ferromagnetic photopolymer was formulated, characterized, and used to fabricate magnetic micropallets, which are microscale pedestals that provide demarcated cell growth surfaces, with preservation of biophysical properties including photopatternability, biocompatibility, and optical clarity. Each micropallet holds a single adherent cell in culture and hundreds of thousands of micropallets compose a single micropallet array. Any micropallet in the array can be recovered on demand, carrying the adhered cell with it. We used this platform to selectively recover single cells, which were subsequently analyzed using single cell RT-qPCR.
The increasing appreciation of tissue cellular heterogeneity and recent identification of rare cell populations within tissues that are associated with specific biological behaviors, e.g., progenitor cells, has illuminated a limitation of current technologies to study such adherent cells directly from primary tissues. The micropallet array is a recently-developed technology designed to address this limitation by virtue of its capacity to isolate and recover single adherent cells on individual micropallets [1]. Micropallet arrays consist of hundreds of thousands of microscale polymer pedestals (“micropallets”) uniformly arrayed on a glass microscope slide. The micropallets are made from a high aspect photopolymerizable polymer using photolithographic methods. Cells are applied to the arrays and fall stochastically upon its surface, with single cells adhering to individual micropallets. Cells are then analyzed in situ and single, unperturbed cells can be selected and collected from the array by releasing the underlying micropallets using a focused pulsed laser.
Rationally designed, individualized therapeutic strategies have long been a desired objective for breast cancer patients and clinicians as an estimated 178,480 new cases of invasive breast cancer will be diagnosed among women in the United States this year and over 40,000 women are expected to die from the disease. [1] The increasing appreciation of breast tumor cellular heterogeneity raises fundamental questions as to the relative contributions of cellular subsets to the biologic behavior of an individual patient’s tumor. [2] As such, it has become increasingly clear that in many cases, an individualized strategy for the treatment of breast cancer would be of great benefit, and that the ability to isolate relevant cellular subsets from the main tumor population is one of the critical limits to accomplishing this goal.
Microelectromechanical systems (MEMS) technologies are ideal for use in sub-millimeter scale actuatable transcatheter optical devices. Such technologies enable precise light path control in very small packages for applications such as optical coherence tomography (OCT) and photodynamic therapies. Indeed, there have been numerous published reports of such devices that utilize silicon-based MEMS technologies and actuation methods including piezoelectric, electrostatic, thermal expansive, and electromagnetic [1–4]. We report a novel, dual axis, magnetically actuated micromirror for endoscopic applications that is fabricated from a photopatternable polymer using photolithographic techniques. Our approach provides improvements over other actuation methods and silicon-based devices.
Light scanning device has many applications and is causing a lot of interest. Optical coherence tomography (OCT) is one example of its application in the field of biomedical imaging. With the trend of miniaturization of biomedical devices, miniaturized light scanning devices are desired for applications like compact OCT system [1].
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