The lysis of cells in order to extract the nucleic acids or proteins inside it is a crucial unit operation in biomolecular analysis. This paper presents a critical evaluation of the various methods that are available both in the macro and micro scale for cell lysis. Various types of cells, the structure of their membranes are discussed initially. Then, various methods that are currently used to lyse cells in the macroscale are discussed and compared. Subsequently, popular methods for micro scale cell lysis and different microfluidic devices used are detailed with their advantages and disadvantages. Finally, a comparison of different techniques used in microfluidics platform has been presented which will be helpful to select method for a particular application.
Specific ranges of
dissolved oxygen (DO) concentrations must be
maintained in a waterbody for it to be hospitable for aquatic animals.
DO sensor designs can employ selectively permeable membranes to isolate
DO from untargeted compounds or organisms in waterbodies. Hence, the
DO concentration can be monitored and the health of the water can
be evaluated over time. However, the presence of bacteria in natural
waterbodies can lead to the formation of biofilms that can block pores
and prevent analyte from permeating the membrane, resulting in inaccurate
readings. In this work, we demonstrate the implementation of a fluorosilane-based
omniphobic lubricant-infused (OLI) coating on a selectively permeable
membrane and investigate the rate of biofilm formation for a commercially
available DO sensor. Coated and unmodified membranes were incubated
in an environment undergoing accelerated bacterial growth, and the
change in sensitivity was evaluated after 40, 100, 250, and 500 h.
Our findings show that the OLI membranes attenuate biofouling by 70%
and maintain sensitivity after 3 weeks of incubation, further demonstrating
that oxygen transfer through the OLI coating is achievable. Meanwhile,
unmodified membranes exhibit significant biofouling that results in
a 3.35 higher rate of decay in oxygen measurement sensitivity and
an over 70% decrease in static contact angle. These results show that
the OLI coating can be applied on commercially available membranes
to prevent biofouling. Therefore, OLI coatings are a suitable candidate
to suppress biofilm formation in the widespread use of selectively
permeable membranes for environmental, medical, and fluid separation
applications.
The incorporation of the extracellular matrix (ECM) is essential for generating in vitro models that truly represent the microarchitecture found in human tissues. However, the cell-cell and cell-ECM interactions in vitro remains poorly understood in placental trophoblast biology. We investigated the effects of varying the surface properties (surface thickness and stiffness) of two ECMs, collagen I and Matrigel, on placental trophoblast cell morphology, viability, proliferation, and expression of markers involved in differentiation/syncytial fusion. Most notably, thicker Matrigel surfaces were found to induce the self-assembly of trophoblast cells into 3D spheroids that exhibited thickness-dependent changes in viability, proliferation, syncytial fusion, and gene expression profiles compared to two-dimensional cultures. Changes in F-actin organization, cell spread morphologies, and integrin and matrix metalloproteinase gene expression profiles, further reveal that the response to surface thickness may be mediated in part through cellular stiffness-sensing mechanisms. Our derivation of self-assembling trophoblast spheroid cultures through regulation of ECM surface alone contributes to a deeper understanding of cell-ECM interactions, and may be important for the advancement of in vitro platforms for research or diagnostics.
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