Rock imager

Manufactured by Formulatrix

Sourced in United States

The Rock Imager is a laboratory instrument designed for automated imaging and analysis of crystallization experiments. It captures high-resolution images of crystal samples at user-defined intervals, providing a comprehensive visual record of the crystallization process.

Automatically generated - may contain errors

Lab products found in correlation

Nt8 drop setter, by Formulatrix (4 mentions) Crystal screen ht, by Hampton Research (3 mentions) Wizard precipitant synergy block no 2, by Molecular Dimensions (3 mentions) Proplex screen ht 96, by Hampton Research (3 mentions) Mosquito crystallization robot, by SPT Labtech (3 mentions) Mosquito lcp robot, by SPT Labtech (2 mentions) Nt8 lcp crystallization robot, by Formulatrix (2 mentions) Monoolein, by Merck Group (2 mentions) Cholesterol, by Merck Group (2 mentions) Lsm 710, by Zeiss (2 mentions) Micromount, by MiTeGen (2 mentions) Mosquito liquid handler, by EQT (1 mentions) Proplex screen, by Molecular Dimensions (1 mentions) Pact premier screen, by Molecular Dimensions (1 mentions) Jcsg plus, by Molecular Dimensions (1 mentions) Morpheus, by Molecular Dimensions (1 mentions) Formulator robot, by Formulatrix (1 mentions) Pact premier, by Molecular Dimensions (1 mentions) Carbon coated copper grid, by Electron Microscopy Sciences (1 mentions) Crystal gryphon, by Art Robbins Instruments (1 mentions)

35 protocols using rock imager

Crystallization of FPR2-WKYMVm Complex

Check if the same lab product or an alternative is used in the 5 most similar protocols

The FPR2-WKYMVm sample was mixed with molten lipid (monoolein and cholesterol 9:1 by mass) at a weight ratio of 1:1.5 (protein:lipid) using two syringes to create a lipidic cubic phase (LCP). The mixture was dispensed onto glass sandwich plates (Shanghai FAstal BioTech) in 40 nl drop and overlaid with 800 nl precipitant solution using a Gryphon robot (Art-Robbins). Protein reconstitution in LCP and crystallization trials were performed at room temperature (19–22 °C). Plates were placed in an incubator (Rock Imager, Formulatrix) and imaged at 20 °C automatically following a schedule. Crystals of FPR2-WKYMVm complex appeared after 4 days and grew to full size (50 × 50 × 5 μm³) within two weeks in 0.1 M Tris, pH 7.0–7.6, 30–36% (v/v) PEG500 DME, 2–5% PPG400, 50–150 mM CH₃COOLi, and 100 μM WKYMVm. The crystals were harvested directly from LCP using 30 and 50 μm micro mounts (M2-L19-30/50, MiTeGen), and flash frozen in liquid nitrogen.

Chen T., Xiong M., Zong X., Ge Y., Zhang H., Wang M., Won Han G., Yi C., Ma L., Ye R.D., Xu Y., Zhao Q, & Wu B. (2020). Structural basis of ligand binding modes at the human formyl peptide receptor 2. Nature Communications, 11, 1208.

+ Open protocol

+ Expand

Crystallization of TCR and TCR-pHLA Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins (TCR or TCR–pHLA complexes) were centrifuged before setting up crystallization trials. The final concentrations ranged between 10 and 20 mg/mL. All proteins were mixed in 96-well Greiner plates either as 2:1, 1:1, or 1:2 ratios, with the screen solution (mother liquor) from commercial primary sparse-matrix screens using a Mosquito liquid handler (SPT Labtech, UK). Final drop volumes were 200 nL; plates were incubated at 20°C in a Rock Imager (Formulatrix, USA). c728 TCR at 19 mg/mL crystallized when mixed 1:2 with 0.2 M lithium sulfate, 0.1 M MES pH 6.0, 20 % w/v PEG 4000 (ProPlex screen, Molecular Dimensions, UK). The following relate to trials using the PACT Premier screen (Molecular Dimensions, UK): c728 TCR–pHLA (at 10.7 mg/mL) crystals grew from 0.02 M sodium/potassium phosphate, 0.1 M bis-Tris propane pH 6.5, 20% w/v PEG 3350; c756 TCR–pHLA (at 10.3 mg/mL) crystals grew from 0.2 M sodium chloride, 0.1 M MES pH 6.0, 20% w/v PEG 6000; c796 TCR–pHLA (at 10.2 mg/mL) crystals grew in 0.2 M ammonium chloride, 0.1 M HEPES pH 7.0, 20% w/v PEG 6000. All crystals were harvested by exchanging first into mother liquor supplemented with 20% glycerol before cryopreservation in liquid nitrogen.
Synchrotron X-ray data were collected with an X-ray wavelength of 0.9763 Å at beamline I03 by Diamond Light Source Industrial Liaison Unit and beamline staff.

Simister P.C., Border E.C., Vieira J.F, & Pumphrey N.J. (2022). Structural insights into engineering a T-cell receptor targeting MAGE-A10 with higher affinity and specificity for cancer immunotherapy. Journal for Immunotherapy of Cancer, 10(7), e004600.

+ Open protocol

+ Expand

Optimizing Protein-Ligand Crystal Structures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Crystallization conditions were screened and monitored with an NT8 drop setter and Rock Imager (Formulatrix). Screening was done with Rigaku Wizard Precipitant Synergy block no. 2 (MD15-PS-B), Molecular Dimensions Proplex screen HT-96 (MD1–38), and Hampton Research Crystal Screen HT (HR2–130) using the sitting drop vapor diffusion method. P-p1f1fab + eOD-GT8 crystals were further optimized with hanging drop trays using vapor diffusion method. Final crystals for P-p1f1fab + eODGT8 were grown in 22.5% PEG 3350, 13.5% Isopropanol, 0.18M Ammonium Citrate pH 4.0. Final crystals for P-p3b3fab + 426c Core were grown in 0.67% PEG 4000, 0.67M Ammonium Citrate pH 5.5. P-p1f1fab + eODGT8 crystal were cryo protected in a solution of 20% molar excess of the crystallization condition and 20% Ethylene Glycol. P-p3b3fab + 426c Core were cryoprotected in the original crystallization condition. P-p3b3fab + 426c Core and P-p1f1fab + eODGT8 were sent to ALS 5.0.2 and diffraction data was collected to 3.59 Å and 3.2 Å respectively. Data were processed using HKL2000 (Otwinowski and Minor, 1997 ).

Parks K.R., MacCamy A.J., Trichka J., Gray M., Weidle C., Borst A.J., Khechaduri A., Takushi B., Agrawal P., Guenaga J., Wyatt R.T., Coler R., Seaman M., LaBranche C., Montefiori D.C., Veesler D., Pancera M., McGuire A, & Stamatatos L. (2019). Overcoming Steric Restrictions of VRC01 HIV-1 Neutralizing Antibodies through Immunization. Cell reports, 29(10), 3060-3072.e7.

+ Open protocol

+ Expand

Protein Reconstitution and Crystallization of P2Y1R

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified protein samples of P2Y₁R were reconstituted into lipidic cubic phase (LCP) by mixing with molten lipid in a mechanical syringe mixer^{33 (link)}. The protein solution was mixed with molten monoolein/cholesterol (10:1 by mass) lipids at weight ratio of 1:1.5 (protein: lipid). After formation of a transparent lipidic cubic phase, the mixture was dispensed onto 96-well glass sandwich plates (Shanghai FAstal BioTech, Inc.) in 40–50 nl drops and overlaid with 800 nl precipitant solution using a Mosquito LCP robot (TTP Labtech). Protein reconstitution in LCP and crystallization trials were performed at room temperature (19–22 °C). Plates were incubated and imaged at 20 °C using an automated incubator/imager (RockImager, Formulatrix). The crystals of the P2Y₁R-MRS2500 complex grew to their full size (70–150 μm) within two weeks in 20–30% PEG400 (v/v), 50–100 mM sodium citrate, 50 μM MRS2500, and 0.1 M HEPES, pH 7.0 or 0.1 M Tris-HCl, pH 8.0. The P2Y₁R-BPTU complex was crystallized in 100–300 mM ammonium phosphate dibasic, 0–10% PEG2000 MME, 50 μM BPTU, and 0.1 M sodium citrate, pH 6.5, and the crystals reached their maximum size (100–130 μm) within two weeks. The P2Y₁R crystals were collected directly from LCP using 50–100 μm micromounts (M2-L19–50/100, MiTeGen) and flash frozen in liquid nitrogen.

Zhang D., Gao Z.G., Zhang K., Kiselev E., Crane S., Wang J., Paoletta S., Yi C., Ma L., Zhang W., Han G.W., Liu H., Cherezov V., Katritch V., Jiang H., Stevens R.C., Jacobson K.A., Zhao Q, & Wu B. (2015). Two disparate ligand binding sites in the human P2Y1 receptor. Nature, 520(7547), 317-321.

+ Open protocol

+ Expand

Optimizing Crystallization Conditions for Calsequestrin

Check if the same lab product or an alternative is used in the 5 most similar protocols

Crystallization screens were carried out in 96-well hanging-drop format and monitored using a Formulatrix Rock Imager automated imaging system. Conditions conducive to crystal growth were optimized and then reproduced in a 24-well format. The best diffraction was obtained by mixing thawed protein (10–20 mg mL^{−1 (link)} in 20 mM HEPES pH 7.3, 400–500 mM NaCl) 1:1 with 15% PEG 4000 and 400 mM Li₂SO₄. The pH of the PEG 4000 solution used to produce the best-diffracting crystals was tested by litmus paper and found to be approximately 3–3.5. Despite the presence of 20 mM HEPES in the protein reagent, the pH of the drops in which crystals grew was controlled by the PEG and remained 3–3.5. Freshly made PEGs were incompatible with calsequestrin crystal growth except when concentrated HCl was added to the mother liquor, producing crystals similar to those observed with benchtop-aged PEGs. Interestingly, only unbuffered conditions yielded crystals. Multiple attempts to grow crystals at a buffered low pH (using acetate or glycine-based buffers) failed.

Titus E.W., Deiter F.H., Shi C., Wojciak J., Scheinman M., Jura N, & Deo R.C. (2020). The structure of a calsequestrin filament reveals mechanisms of familial arrhythmia. Nature structural & molecular biology, 27(12), 1142-1151.

+ Open protocol

+ Expand

Optimized Protein Crystallization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein crystallization was performed at the Division of Structural Biology of the Wellcome Centre for Human Genetics, University of Oxford. Initial screening was performed using the sitting drop method in 300 nL (100 nL of protein solution and 200 nL of crystallization reagents) using the Hydra and Cartesian instruments. All crystallization plates were stored in Rock Imager (Formulatrix, Bedford, MA, USA) at 21 °C. Four commercially available screens (Index, Proplex, PACTpremier, Crystal screen) were used for the equimolar mixture of deglycosylated B7-H6 with NKp30_Stalk or NKp30_LBD, both expressed in HEK293S GnTI⁻ cells; the mixture was concentrated to 10 mg/mL before the drop set-up. Initially, needle-shaped crystals of NKp30_LBD:B7-H6 complex were obtained in 0.1 M sodium citrate pH 5.0, 20% PEG 8000. These crystals were crushed and used for seeding in optimization performed using the sitting drop method in 300 nL (200 nL of protein solution and 100 nL of crystallization reagents). The number of crystals grew in the drops that were seeded, and diffraction data were collected for a few of them. 25% glycerol was used as cryoprotectant and data were collected at 100 K. The best diffraction data were collected from crystals grown in 0.1 M sodium citrate pH 6.7, 11.7% PEG 6000.

Skořepa O., Pazicky S., Kalousková B., Bláha J., Abreu C., Ječmen T., Rosůlek M., Fish A., Sedivy A., Harlos K., Dohnálek J., Skálová T, & Vaněk O. (2020). Natural Killer Cell Activation Receptor NKp30 Oligomerization Depends on Its N-Glycosylation. Cancers, 12(7), 1998.

+ Open protocol

+ Expand

Vapour Diffusion Crystallization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

All vapour diffusion crystallization experiments were performed using 96-well, 2-drop ‘MRC’ plates. For each protein, crystallization screening trials were performed with the JCSG Plus, PACT Premier and Morpheus screens (Molecular Dimensions), using 0.1 µL crystallization buffer mixed with 0.1 μL protein (7.5 mg/mL), both with and without 2–5 mM hmdUMP (Jena Bioscience or Biosynth Carbosynth), in separate drop positions. Where necessary, optimisation screens based on hits in the presence of substrate were subsequently generated using a Formulatrix Formulator robot and dispensed as before except using protein at 5 and 7.5 mg/mL, both in the presence of 5 mM substrate. All trays were dispensed at room temperature using an NT8 humidity-controlled robot (Formulatrix) but stored at 4 °C and automatically imaged by a Formulatrix Rock Imager.
Crystal harvesting was performed at 4 °C, with crystals cryo-protected stepwise using varying ratios of crystallization buffer:cryoprotectant (2:1, 1:2, 0:1). See Supplementary Table 3 for optimised crystallization conditions and respective cryoprotectants.

Rzechorzek N.J., Kunzelmann S., Purkiss A.G., Silva Dos Santos M., MacRae J.I., Taylor I.A., Fugger K, & West S.C. (2023). Mechanism of substrate hydrolysis by the human nucleotide pool sanitiser DNPH1. Nature Communications, 14, 6809.

+ Open protocol

+ Expand

Crystallization and Structure Determination of Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols

Crystallization conditions were screened and monitored with an NT8 drop setter and Rock Imager (Formulatrix). Screening was done with Rigaku Wizard Precipitant Synergy block no. 2, Molecular Dimensions Proplex screen HT-96, and Hampton Research Crystal Screen HT using the sitting drop vapor diffusion method. Final crystals were grown in a solution of 18.43% PEG 3350, 11- .01%, Lithium Sulfate, 0.11 M Imidazole pH 6.5. Crystals were cryoprotected in a solution of 30% excess of the crystallization condition and 20% glycerol. Crystals were sent to ALS 5.0.1 and diffraction data was collected to 3.42 Å. Data was processed using HKL-2000 (Otwinowski and Minor, 1997 ). The structure was solved using molecular replacement using PDB ID: 5IFA as a search model in Phaser in Phenix (Adams et al., 2010 (link)). The structure was further refined with COOT (Emsley and Cowtan, 2004 (link)) and Phenix (Adams et al., 2010 (link)). The refinement statistics are summarized in Supplemental Table 2.

Lin Y.R., Rachael Parks K., Weidle C., Naidu A.S., Khechaduri A., Riker A.O., Takushi B., Chun J.H., Borst A.J., Veesler D., Stuart A., Agrawal P., Gray M., Pancera M., Huang P.S, & Stamatatos L. (2020). HIV-1 VRC01 germline-targeting immunogens select distinct epitope-specific B cell receptors. Immunity, 53(4), 840-851.e6.

+ Open protocol

+ Expand

Characterizing Tofacitinib Crystals via Microscopy

Check if the same lab product or an alternative is used in the 5 most similar protocols

For Confocal Microscopy, 10 µL of the MTH was placed on a glass slide, covered with a cover slip and sealed with nail polish. Differential Interference Contrast (DIC) channel of a confocal microscope (Zeiss LSM 710) was used to characterize the tofacitinib crystals suspended in the MTH matrix as well as the crystals formed in the absence of the MTH hydrogel.
For Polarized Light Microscopy, MTH samples were dispensed into the sample well of a crystallization plate (Swissci/MRC UV-XPO) and imaged using a Rock Imager (Formulatrix) by cross-polarized visible light. Second Harmonic Generation (SHG) images were also acquired similarly.
For Transmission Electron Microscopy (TEM), MTH samples were diluted 100X with water and 5 µL the solution was applied to 200 mesh carbon-coated copper grids (Electron Microscopy Sciences). The sample was applied for 30 s and blotted with a filter paper. Aqueous uranyl formate (0.75%, w/v) was used for negative staining. Images of the fibril nanostructure were acquired on a Hitachi 7650 transmission electron microscope at 80 kv accelerating voltage.

Majumder P., Zhang Y., Iglesias M., Fan L., Kelley J.A., Andrews C., Patel N., Stagno J.R., Oh B.C., Furtmüller G.J., Lai C.C., Wang Y.X., Brandacher G., Raimondi G, & Schneider J.P. (2020). Multiphase Assembly of Small Molecule Microcrystalline Peptide Hydrogel Allows Immunomodulatory Combination Therapy for Long-term Heart Transplant Survival. Small (Weinheim an der Bergstrasse, Germany), 16(38), e2002791.

+ Open protocol

+ Expand

Crystallization of CD47 Complex

Check if the same lab product or an alternative is used in the 5 most similar protocols

The CD47 complex samples were mixed with molten lipid (monoolein and cholesterol at a 9:1 ratio by mass) at a weight ratio of 1:1.5 (protein:lipid) using two syringes to create a LCP^{24 (link)}. The mixture was dispensed onto glass sandwich plates in 40 nl drop and overlaid with 800 nl precipitant solution using a MOSQUITO LCP robot (labtech). Protein reconstitution in LCP and crystallization trials were performed at room temperature (19–22 °C). Plates were placed in an incubator (Rock Imager, Formulatrix) and imaged at 20 °C automatically following a schedule. Crystals of the CD47^BRIL-B6H12 complex appeared in 48–72 h and grew to a full size of ~55 × 50 × 15 μm³ within 4 weeks in 0.1 M sodium citrate pH 5.9–6.2, 450–500 mM ammonium acetate, and 32–35% PEG400. The crystals were harvested directly from LCP using 30 and 50 μm micro mounts (M2-L19-30/50, MiTeGen), and flash-cooled in liquid nitrogen.

Fenalti G., Villanueva N., Griffith M., Pagarigan B., Lakkaraju S.K., Huang R.Y., Ladygina N., Sharma A., Mikolon D., Abbasian M., Johnson J., Hadjivassiliou H., Zhu D., Chamberlain P.P., Cho H, & Hariharan K. (2021). Structure of the human marker of self 5-transmembrane receptor CD47. Nature Communications, 12, 5218.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!