scholarly journals Toward the Precise Control of Cell Differentiation Processes by Using Micro and Soft Lithography

Author(s):  
Yuzo Takayama ◽  
Hiroyuki Moriguchi ◽  
Kiyoshi Kotani ◽  
Takafumi Suzuki ◽  
Kunihiko Mabuchi ◽  
...  
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Hye Ji Cha ◽  
Özgün Uyan ◽  
Yan Kai ◽  
Tianxin Liu ◽  
Qian Zhu ◽  
...  

AbstractPrecise control of gene expression during differentiation relies on the interplay of chromatin and nuclear structure. Despite an established contribution of nuclear membrane proteins to developmental gene regulation, little is known regarding the role of inner nuclear proteins. Here we demonstrate that loss of the nuclear scaffolding protein Matrin-3 (Matr3) in erythroid cells leads to morphological and gene expression changes characteristic of accelerated maturation, as well as broad alterations in chromatin organization similar to those accompanying differentiation. Matr3 protein interacts with CTCF and the cohesin complex, and its loss perturbs their occupancy at a subset of sites. Destabilization of CTCF and cohesin binding correlates with altered transcription and accelerated differentiation. This association is conserved in embryonic stem cells. Our findings indicate Matr3 negatively affects cell fate transitions and demonstrate that a critical inner nuclear protein impacts occupancy of architectural factors, culminating in broad effects on chromatin organization and cell differentiation.


2018 ◽  
Author(s):  
Andre Lai ◽  
Nicolas Altemose ◽  
Jonathan A. White ◽  
Aaron M. Streets

AbstractIntegrated elastomeric valves, also referred to as Quake valves, enable precise control and manipulation of fluid within microfluidic devices. Fabrication of such valves requires bonding of multiple layers of the silicone polymer polydimethylsiloxane (PDMS). The conventional method for PDMS-PDMS bonding is to use varied base to crosslinking agent ratios between layers, typically 20:1 and 5:1. This bonding technique, known as “off-ratio bonding,” provides strong, effective PDMS-PDMS bonding for multi-layer soft-lithography, but it can yield adverse PDMS material properties and can be wasteful of PDMS. Here we demonstrate the effectiveness of on-ratio PDMS bonding for multilayer soft lithography. We show the efficacy of this technique among common variants of PDMS: Sylgard 184, RTV 615, and Sylgard 182.


Materials ◽  
2021 ◽  
Vol 14 (23) ◽  
pp. 7275
Author(s):  
Koungjun Min ◽  
Jaemook Lim ◽  
Ji Hwan Lim ◽  
Eunseung Hwang ◽  
Youngchan Kim ◽  
...  

Poly(dimethylsiloxane) has attracted much attention in soft lithography and has also been preferred as a platform for a photochemical reaction, thanks to its outstanding characteristics including ease of use, nontoxicity, and high optical transmittance. However, the low stiffness of PDMS, an obvious advantage for soft lithography, is often treated as an obstacle in conducting precise handling or maintaining its structural integrity. For these reasons, a Glass-PDMS-Glass structure has emerged as a straightforward alternative. Nevertheless, several challenges are remaining in fabricating Glass-PDMS-Glass structure through the conventional PDMS patterning techniques such as photolithography and etching processes for master mold. The complicated techniques are not suitable for frequent design modifications in research-oriented fields, and fabrication of perforated PDMS is hard to achieve using mold replication. Herein, we utilize the successive laser pyrolysis technique to pattern thin-film PDMS for microfluidic applications. The direct use of thin film at the glass surface prevents the difficulties of thin-film handling. Through the precise control of photothermal pyrolysis phenomena, we provide a facile fabrication process for perforated PDMS microchannels. In the final demonstration, the laminar flow has been successfully created owing to the smooth surface profile. We envision further applications using rapid prototyping of the perforated PDMS microchannel.


Author(s):  
A. Engel ◽  
A. Holzenburg ◽  
K. Stauffer ◽  
J. Rosenbusch ◽  
U. Aebi

Reconstitution of solubilized and purified membrane proteins in the presence of phospholipids into vesicles allows their functions to be studied by simple bulk measurements (e.g. diffusion of differently sized solutes) or by conductance measurements after transformation into planar membranes. On the other hand, reconstitution into regular protein-lipid arrays, usually forming at a specific lipid-to-protein ratio, provides the basis for determining the 3-dimensional structure of membrane proteins employing the tools of electron crystallography.To refine reconstitution conditions for reproducibly inducing formation of large and highly ordered protein-lipid membranes that are suitable for both electron crystallography and patch clamping experiments aimed at their functional characterization, we built a flow-dialysis device that allows precise control of temperature and flow-rate (Fig. 1). The flow rate is generated by a peristaltic pump and can be adjusted from 1 to 500 ml/h. The dialysis buffer is brought to a preselected temperature during its travel through a meandering path before it enters the dialysis reservoir. A Z-80 based computer controls a Peltier element allowing the temperature profile to be programmed as function of time.


Author(s):  
M.V. Parthasarathy ◽  
C. Daugherty

The versatility of Low Temperature Field Emission SEM (LTFESEM) for viewing frozen-hydrated biological specimens, and the high resolutions that can be obtained with such instruments have been well documented. Studies done with LTFESEM have been usually limited to the viewing of small organisms, organs, cells, and organelles, or viewing such specimens after fracturing them.We use a Hitachi 4500 FESEM equipped with a recently developed BAL-TEC SCE 020 cryopreparation/transfer device for our LTFESEM studies. The SCE 020 is similar in design to the older SCU 020 except that instead of having a dedicated stage, the SCE 020 has a detachable cold stage that mounts on to the FESEM stage when needed. Since the SCE 020 has a precisely controlled lock manipulator for transferring the specimen table from the cryopreparation chamber to the cold stage in the FESEM, and also has a motor driven microtome for precise control of specimen fracture, we have explored the feasibility of using the LTFESEM for multiple-fracture studies of the same sample.


Author(s):  
H. Alasam

The possibility that intrathymic T-cell differentiation involves stem cell-lymphoid interactions in embryos led us to study the ultrastructure of epithelial cell in normal embryonic thymus. Studies in adult thymus showed that it produces several peptides that induce T-cell differentiation. Several of them have been chemically characterized, such as thymosin α 1, thymopoietin, thymic humoral factor or the serum thymic factor. It was suggested that most of these factors are secreted by populations of A and B-epithelial cells.Embryonic materials were obtained from inbred matings of Swiss Albino mice. Thymuses were disected from embryos 17 days old and prepared for transmission electron microscopy. Our studies showed that embryonic thymus at this stage contains undifferentiated and differentiated epithelial cells, large lymphoblasts, medium and few small lymphocytes (Fig. 5). No differences were found between cortical and medullary epithelial cells, in contrast to the findings of Van Vliet et al,. Epithelial cells were mostly of the A-type with low electron density in both cytoplasm and nucleus. However few B-type with high electron density were also found (Fig. 7).


2001 ◽  
Vol 120 (5) ◽  
pp. A517-A517
Author(s):  
A MIZOGUCHI ◽  
E MIZOGUCHI ◽  
Y DEJONG ◽  
H TAKEDATSU ◽  
F PREFFER ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document