手动压力生成器
Overview of 高压单元系统
高压元件 (HP元件) 系统包括了压力单元、泵以及用于系统控制和数据获取的软件。它专为固体和液体样本的吸收、荧光、拉曼光谱和旋光研究而设计。该元件由不锈钢合金制成，用静液压泵可以达到上至400 MPa (4,000巴) 的压力。这款HP元件可以被直接安装在ISS的分光荧光计和多种商用仪器中，同时它也可以作为一个独立单元使用。
这款HP元件有3窗或4窗版本，各个窗口片之间呈90度。石英和蓝宝石窗口片拥有一个10 mm的小孔；氟化镁窗口片 (用于研究旋光) 有一个3 mm的小孔。
拥有10 mm高孔径的窗口片可以使用激光器、发光二极管 (LED) 或来自氙弧灯的准直光束作为激发光源。对于这样的元件，它格外高的数值孔径 (NA = 0.18) 使其在需要高灵敏度的荧光研究中独一无二。
与之兼容的仪器： ChronosBH, ChronosDFD, and PC1.电贯通件
每当需要对样品施加电压时，HP单元都可以使用插头。插头被装到HP单元的顶部；通过将弹性管从泵水平连接到插头上可以施加压力。使用插头可以达到上至300 MPa的压力
HP单元带有用于电贯通件的插头。压力通过水平连接到插头的管道施加。电压被施加在电极上。
窗口
光学窗口是由石英石 (SiO2)、蓝宝石 (单晶氧化铝 Al2O3) 或者MgF2制成的。窗口被安装到抛光的插头上，它们由精准的机械支架固定
| 材料 | 最大压力 (MPa) | 孔径 (mm) | 双折射 (Δn) | 传播范围 (nm) |
|---|---|---|---|---|
| 石英 [SiO2] | 300 | 10 | +0.009 | 180 - 2,000 |
| 蓝宝石 [Al2O3] | 400 | 10 | -0.008 | 250 - 5,000 |
| MgF2 | 250 | 5 | +0.006 | 150 - 6,500 |
窗口材料由具体应用和要获取的测量结果决定。石英窗是由UV级熔融石英制成的，它可以承受上至300 MPa的压强。熔融石英是一种多晶、各向同性、且没有晶体取向的材料；这些窗口适合用于偏振测量。蓝宝石窗口能够承受上至400 MPa的压强。但是石英是一种合成的六方单晶各向异性材料，它在沿不同轴测量时会表现出截然不同的光学特征。如果蓝宝石是双折射的，也就是沿着光轴的光束会经历两个不同的折射率值。这些窗口没有那么适合用于测量偏振。MgF2窗口拥有最小的双折射； 此外，它们的传输扩展到了150 nm，这使得它们适合用于圆二色性研究。
温度控制
这款高压单元包括了一个内置路径，用于从直接与其连接的外部浴循环器对温度控制液体进行循环。由于该单元是由导热性优异的不锈钢合金制成的，稳定温度的过程十分迅速。这款高压单元设计用于在-40 °C至80 °C的温度范围内运作。
Product Specifications for 高压单元系统
细胞
压力
- 上至300 MPa (3,000巴) 的压力，带有石英窗
- 上至400 MPa (4,000巴) 的压力，带有蓝宝石窗
- 上至250 MPa (2,500巴) 的压力，带有MgF2窗
温度
- -40 °C to +80 °C
- 通过内置液体循环回路进行控制
- 导热系数
- 2 W • cm-1 • K-1 at 20 °C
- 16 W • cm-1 • K-1 at 200 °C
窗口
- 石英 (直径19 mm粗细 x 8.5 mm)
- 蓝宝石 (直径19 mm粗细 x 6.4 mm)
- MgF2 (直径19 mm粗细 x 8.5 mm)
窗口小孔
- 石英石和蓝宝石：10 mm
- MgF2: 3 mm
高压单元内室
- 直径：22 mm
- 高度：50.3 mm
大小
- 119 mm (宽) x 105 mm (长) x 121.5 mm (高)
- 从底部到窗口中心为39 mm
压力生成器
测量
- 上至600 MPa
储液量
- 116 ml
生成器体积
- 10 ml (手动或自动泵)
管道
- 直径1/8" ，柔性上至415 MPa
热电偶
- Ni-Cr-Ni或是Fe-Const
样品支架体积
空单元，3个石英窗
- 15.0 ml
空单元，4个蓝宝石窗
- 24.5 ml
支架被配有11 mm的瓶和盖
- 11.0 ml
- 瓶和盖加起来的最大允许高度：40.0 mm
圆形比色皿，11 mm外直径
- 0.75 ml
圆形比色皿，9mm外直径
- 0.80 ml
方形单元，6 x 6 mm
- 0.30 ml
静压液体
压力媒介
- 光谱级水
- 低荧光背景乙醇
高压单元系统的产品配件
产品资源
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“Response and adaptation of the transcriptional heat shock response to pressure.” Coffin, C.H., Fisher, L.A., Crippen, S., Demers, P., Bartlett, D.H., & Royer, C.A. Frontiers in Microbiology, 15, 2024, Nov. doi: 10.3389/fmicb.2024.1470617. -
“Ions in the Deep Subsurface of Earth, Mars, and Icy Moons: Their Effects in Combination with Temperature and Pressure on tRNA–Ligand Binding.” Jahmidi-Azizi, N., Gault, S., Cockell, C.S., Oliva, R., & Winter, R. International Journal of Molecular Sciences, 22(19), 2021, Oct. doi: 10.3390/ijms221910861.
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“The consequences of cavity creation on the folding landscape of a repeat protein depend upon context.” Jenkins, K.A., Fossat, M.J., Zhang, S., Rai, D.K., Klein, S., Gillilan, R., White, Z., Gerlich, G., McCallum, S.A., Winter, R., Gruner, S.M., Barrick, D., & Royer, C.A. PNAS, 115(35), pp. E8153–E8161, 2018, Aug. doi: 10.1073/pnas.1807379115.
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“Targeting the Prion-like Aggregation of Mutant p53 to Combat Cancer.” Silva, J.L., Cino, E.A., Soares, L.N., Ferreira, V.F., & de Oliveira, A.P. Accounts of Chemical Research, 51(1), pp. 181–190, 2017, Dec. doi: 10.1021/acs.accounts.7b00473.
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“Comparing Fast Pressure Jump and Temperature Jump Protein Folding Experiments and Simulations.” Wirth, A.J., Liu, Y., Prigozhin, M.B., Schulten, K. & Gruebele, M. Journal of the American Chemical Society, 137(22), pp. 7152–7159, 2015, Jun. doi: 10.1021/jacs.5b02474.
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“Dual chamber shape memory alloy unplugging and mixing system coupled to a high pressure optical cell for biophysical studies.” Leist, S.K., Kunkle, C., Chong, P.L.G. & Zhou, J. Smart Materials and Structures, 24(3), 2015, Feb. doi: 10.1088/0964-1726/24/3/035014.
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“Circular dichroism and site-directed spin labeling reveal structural and dynamical features of high-pressure states of myoglobin.” Lerch, M.T., Horwitz, J., Mccoy, J. & Hubbell, W.L. Proceedings of the National Academy of Sciences, 110(49), 2013, Nov. doi: 10.1073/pnas.1320124110.
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“Pressure Modulation of Ras–Membrane Interactions and Intervesicle Transfer.” Kapoor, S., Werkmüller, A., Goody, R.S., Waldmann, H. & Winter, R. Journal of the American Chemical Society, 135(16), pp. 6149–6156, 2013, Apr. doi: 10.1021/ja312671j.
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“Misplaced helix slows down ultrafast pressure-jump protein folding.” Prigozhin, M.B., Liu, Y., Wirth, A.J., Kapoor, S., Winter, R., Schulten, K. & Gruebele, M. Proceedings of the National Academy of Sciences, 110(20), pp. 8087–8092, 2013, Apr. doi: 10.1073/pnas.1219163110.
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“Dissecting the Structure, Thermodynamic Stability, and Aggregation Properties of the A25T Transthyretin (A25T-TTR) Variant Involved in Leptomeningeal Amyloidosis: Identifying Protein Partners That Co-Aggregate during A25T-TTR Fibrillogenesis in Cerebrospinal Fluid.” Azevedo, E.P.C., Pereira, H.M., Garratt, R.C., Kelly, J.W., Foguel, D. & Palhano, F.L. Biochemistry, 50(51), pp. 11070–11083, 2011, Nov. doi: 10.1021/bi201365r.
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“Refolding of endostatin from inclusion bodies using high hydrostatic pressure.” Chura-Chambi, R.M., Genova, L.A., Affonso, R. & Morganti, L. Analytical Biochemistry, 379(1), pp. 32–39, 2008, Aug. doi: 10.1016/j.ab.2008.04.024.
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“Vi -Value Analysis: A Pressure-Based Method for Mapping the Folding Transition State Ensemble of Proteins.” Mitra, L., Hata, K., Kono, R., Maeno, A., Isom, D., Rouget, J.-B., Winter, R., Akasaka, K., García-Moreno, B. & Royer, C.A. Journal of the American Chemical Society, 129(46), pp. 14108–14109, 2007, Oct. doi: 10.1021/ja073576y.
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“pH dependence of the dissociation of multimeric hemoglobin probed by high hydrostatic pressure.” Bispo, J.A., Santos, J.L., Landini, G.F., Goncalves, J.M. & Bonafe, C.F. Biophysical Chemistry, 125(2-3), pp. 341–349, 2007, Feb. doi: 10.1016/j.bpc.2006.09.009.
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“Proton dependence of tobacco mosaic virus dissociation by pressure.” Santos, J.L., Bispo, J.A., Landini, G.F. & Bonafe, C.F. Biophysical Chemistry, 111(1), pp. 53–61, 2004, Sep. doi: 10.1016/j.bpc.2004.04.003.
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“Pressure and temperature dependence of the melt segmental dynamics of cis-1,4-polyisoprene via time resolved optical spectroscopy.” Punchard, B.J. & Adolf, D.B. The Journal of Chemical Physics, 117(16), pp. 7774–7780, 2002, Oct. doi: 10.1063/1.1509450.
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“Effects of Pressure-Induced Membrane Phase Transitions on Inactivation of HorA, an ATP-Dependent Multidrug Resistance Transporter, in Lactobacillus plantarum.” Ulmer, H.M., Herberhold, H., Fahsel, S., Gänzle, M.G., Winter, R., & Vogel, R.F. Applied and Environmental Microbiology, 68(3), pp. 1088–1095, 2002, Mar. doi: 10.1128/AEM.68.3.1088–1095.2002.
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“Pressure and Temperature Dependence of the Dilute Solution Segmental Dynamics of Anthracene-Labeled Polyisoprene.” Punchard, B.J. & Adolf, D.B. Macromolecules, 35(8), pp. 3281–3287, 2002, Mar. doi: 10.1021/ma011783r.
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“Time-resolved optical spectroscopy study of the local dynamics of cis-1,4 and vinyl-1,2-polybutadiene in dilute solution at high pressure.” Punchard, B., Kirpatch, A. & Adolf, D. Polymer, 43(23), pp. 6287–6293, 2002, Jan. doi: 10.1016/s0032-3861(02)00549-9.
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“Unusual Properties of Highly Charged Buffers: Large Ionization Volumes and Low Barrier Hydrogen Bonds.” Hess, R.A. & Reinhardt, L.A. Journal of the American Chemical Society, 121(42), pp. 9867–9870, 1999, Oct. doi: 10.1021/ja9921726.
兼容性
ISS提供用于将高压单元安装至各个荧光分光计和分光光度计的合适底盘。如果高压单元不能被装到仪器上，光纤束可以被用于连接。请联系ISS以了解更多有关光纤套件的细节。
荧光分光计安装套件可被用于
| ISS | PC1 K2 ChronosBH ChronosDFD |
|---|---|
| AVIV | AFTF105 |
| Edinburgh仪器 | FS900 |
| Hitachi | F4500 |
| Horiba | Fluorolog 2, 3 Fluoromax 2, 3 |
| Jasco | FP-8300, FP-8500, FP-8600 |
| SLM | 4800, 8000, 8100, 48000 |
| Varian | Cary Eclipse |
分光光度计套件可用于
| Jasco | V-750, V-760, V-770, V-780 |
|---|---|
| Shimadzu | UV2501PC, UV1650PC |
| Agilent | Cary 5000 |
分光旋光计安装套件可用于：
| Jasco | J-710, J-715, J-810, J-815, J-1500, J-1700 |
|---|
ISS PC1荧光分光计中的高压单元
ISS ChronosDFD寿命荧光分光计中的高压单元
靠近ISS ChronosDFD的手动泵
连接至压力泵的高压单元
SLM 8100型号荧光分光计中的高压单元
Jasco 8300型号荧光分光计中的高压单元
Varian Cary100分光光度计中的高压单元
寿命测量
(由蒙彼利埃大学 (法国蒙彼利埃) 生物结构中心的Catherine A. Royer教授提供)
吸收和发射光谱
(承蒙美国宾州费城天普大学Parkson L.-G. Chong教授的支持)
