Article
Materials Science, Multidisciplinary
Matthew Lindley, Julia Gala de Pablo, Jorgen Walker Peterson, Akihiro Isozaki, Kotaro Hiramatsu, Keisuke Goda
Summary: Cell sorting is an important tool in biological research and medicine. Raman-activated cell sorting (RACS) has gained attention for its ability to discriminate cells based on their intracellular chemical content without the need for labeling. However, the widespread use of RACS has been limited by a trade-off between throughput and measurement bandwidth. In this study, this trade-off is overcome and broadband RACS in the fingerprint region with a record high throughput is demonstrated. Real-time label-free sorting of microalgal cells based on their accumulation of carotenoids and polysaccharide granules is also showcased.
ADVANCED MATERIALS TECHNOLOGIES
(2022)
Article
Biotechnology & Applied Microbiology
Ashley E. Sproles, Anthony Berndt, Francis J. Fields, Stephen P. Mayfield
Summary: This study describes the design of an optimized vector for rapid identification and isolation of high-expression recombinant protein transformants in Chlamydomonas. The method greatly reduces screening time and enables the production of large populations of recombinant algae transformants for subsequent sequencing and analysis.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Analytical
Alex A. Zhukov, Robyn H. Pritchard, Mick J. Withers, Tony Hailes, Richard D. Gold, Calum Hayes, Mette F. la Cour, Fred Hussein, Salman Samson Rogers
Summary: By creating a parallel version of the vortex-actuated cell sorter (VACS), researchers achieved high-throughput microfluidic cell sorting. This approach sorts lymphocyte-sized particles at a rate 16 times faster than single-stream VACS devices and 10 times faster than commercial cell sorters, offering potential for processing much greater cell numbers in applications.
Review
Biotechnology & Applied Microbiology
Jingjie Jiang, Guangyu Yang, Fuqiang Ma
Summary: Fluorescence-activated droplet sorting (FADS) is a powerful tool for high-throughput screening of enzymes, metabolites, and antibodies. Fluorescence coupling strategies (FCSs) play a key role in developing new FADS methods by linking analyte properties to fluorescence signals. This review highlights recent advances in FCS for different analyte types and discusses successful FADS applications.
BIOTECHNOLOGY ADVANCES
(2023)
Article
Biochemistry & Molecular Biology
Gheorghita Menghiu, Vasile Ostafe, Radivoje Prodanovic, Rainer Fischer, Raluca Ostafe
Summary: Chitinases catalyze the degradation of chitin, a polymer found in various organisms. Developing improved chitinases is important for addressing environmental concerns and potential applications of chitosan products. This study focused on creating a high-throughput screening system and selecting enhanced chitinase variants by utilizing fluorescence-activated cell sorting and mutagenesis techniques.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Cell Biology
Xuan Xie, Tomas Tichopad, Galina Kislik, Lucie Langerova, Pavel Abaffy, Radek Sindelka, Roman Franek, Michaela ikova, Christoph Steinbach, Mujahid Ali Shah, Ivo Sauman, Fan Chen, Martin Psenicka
Summary: The study successfully identified and isolated type A spermatogonia from different developmental stages of testes using flow cytometric cell sorting (FCM), with the P1 gate containing the highest amount of type A spermatogonia and the P2 gate mainly containing later maturing type B spermatogonia. Additionally, genes such as plzf were found to have significantly higher expressions in type A spermatogonia, providing potential for improving efficiency in germ stem cell culture and transplantation.
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Takeshi Tabuchi, Yohei Yokobayashi
Summary: Cell-free systems without living cells are promising platforms for testing and practical applications. However, biological constraints limit the development of cell-free devices using living cells. This study presents a high-throughput screening method for engineering cell-free riboswitches that respond to small molecules.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Multidisciplinary Sciences
Jongmin Woo, Sarah M. Williams, Lye Meng Markillie, Song Feng, Chai-Feng Tsai, Victor Aguilera-Vazquez, Ryan L. Sontag, Ronald J. Moore, Dehong Hu, Hardeep S. Mehta, Joshua Cantlon-Bruce, Tao Liu, Joshua N. Adkins, Richard D. Smith, Geremy C. Clair, Ljiljana Pasa-Tolic, Ying Zhu
Summary: This study introduces a nested nanoPOTS (N2) chip to improve protein recovery, operation robustness, and processing throughput for isobaric-labeling-based single-cell proteomics workflow. The N2 chip enables robust quantification of approximately 1500 proteins and reveals membrane protein markers, demonstrating high stability of single-cell proteome profiles for cells cultured under identical conditions.
NATURE COMMUNICATIONS
(2021)
Article
Chemistry, Analytical
Gerjen H. Tinnevelt, Kristiaan Wouters, Geert J. Postma, Rita Folcarelli, Jeroen J. Jansen
Summary: White blood cells play a crucial role in protecting the body from diseases, but can also be related to chronic inflammation, auto immune diseases, or leukemia. High-throughput analytical instruments have been developed to measure multiple proteins on millions of single cells to study the identity and function of different white blood cell types. Multivariate statistics are essential for fully extracting the information-rich biochemistry data. The process involves analyzing the study design, formulating a research question, preparing the data, converting single cells into a cellular distribution, and using clustering methods or models for analysis to differentiate cell (sub)types between groups.
ANALYTICA CHIMICA ACTA
(2021)
Article
Biochemical Research Methods
Jessica T. Stieglitz, James A. Van Deventer
Summary: This study developed a yeast display-based platform to screen for previously undiscovered variants of aminoacyl-tRNA synthetases (aaRSs). The findings expand the options for genetic code manipulation in yeast and have important implications for the fundamental properties and evolvability of aaRSs. Access to diverse orthogonal translation systems is invaluable for applications in chemical biology, synthetic biology, and protein engineering.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Biochemical Research Methods
Jessica T. Stieglitz, James A. Van Deventer
Summary: Protein expression with genetically encoded noncanonical amino acids (ncAAs) has various applications, but the lack of orthogonal translation systems (OTSs) limits their use. This study uses yeast display and flow cytometry to screen libraries of aaRSs for incorporating new ncAAs, improving existing aaRSs, and creating highly selective OTSs. The discovery of previously unknown aaRS variants significantly increases the available options for genetic code manipulation in yeast.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Biochemical Research Methods
Xiaoguang Lu, Joycelyn Jia Ming Chow, Seok Hwee Koo, Boran Jiang, Thean Yen Tan, Dahou Yang, Ye Ai
Summary: A novel inertial microfluidic method has been developed to effectively sort bacteria from larger blood cells, improving pathogen detection efficiency and accuracy.
Article
Optics
B. Ardini, A. BAssI, A. Candeo, A. Genco, C. Trovatello, F. Liu, X. Zhu, G. Valentini, G. Cerullo, R. Vanna, C. Manzoni
Summary: Raman microscopy is a powerful technique for mapping the chemical composition of a sample. This paper presents a compact wide-field Raman microscope based on a time-domain Fourier-transform approach, allowing for parallel acquisition of Raman spectra on all pixels of a 2D detector. The system has high spatial and spectral resolutions and the ability to separate fluorescence and Raman signals.
Article
Chemistry, Analytical
Andrew P. Burns, Ya-Qin Zhang, Tuan Xu, Zhengxi Wei, Qin Yao, Yuhong Fang, Valeriu Cebotaru, Menghang Xia, Matthew D. Hall, Ruili Huang, Anton Simeonov, Christopher A. LeClair, Dingyin Tao
Summary: The development of a highly automated universal sample preparation platform has significantly improved the efficiency of proteomics sample preparation, enabling high-throughput extraction and analysis of proteins from various human cell types. This platform demonstrates excellent digestion efficiency, reproducibility, and universality in quantifying target proteins, showcasing its potential for clinical research.
ANALYTICAL CHEMISTRY
(2021)
Article
Chemistry, Multidisciplinary
Ngoc Duc Vo, Anh Thi Van Nguyen, Hoi Thi Le, Nam Hoang Nguyen, Huong Thi Thu Pham
Summary: The study developed an alternative method, the MACS-Countess system, for CD4(+) T cell counting, which combines magnetic activated cell sorting with an automated cell counting system. Compared to the FACS gold standard, the MACS-Countess system showed a mean bias of 3.5% with a narrow limit of agreement, making it a reliable and cost-effective option for laboratories in limited resource regions.
APPLIED SCIENCES-BASEL
(2021)
Article
Hematology
Andres C. Garcia-Montero, Maria Jara-Acevedo, Ivan Alvarez-Twose, Cristina Teodosio, Laura Sanchez-Munoz, Carmen Muniz, Javier I. Munoz-Gonzalez, Andrea Mayado, Almudena Matito, Carolina Caldas, Jose M. Morgado, Luis Escribano, Alberto Orfao
Article
Hematology
Laura Sanchez-Munoz, Jose M. Morgado, Ivan Alvarez-Twose, Almudena Matito, Andres C. Garcia-Montero, Cristina Teodosio, Maria Jara-Acevedo, Andrea Mayado, Manuela Mollejo, Carolina Caldas, David Gonzalez de Olano, Luis Escribano, Alberto Orfao
BRITISH JOURNAL OF HAEMATOLOGY
(2016)
Article
Allergy
Ivan Alvarez-Twose, Maria Jara-Acevedo, Jose Mario Morgado, Andres Garcia-Montero, Laura Sanchez-Munoz, Cristina Teodosio, Almudena Matito, Andrea Mayado, Carolina Caldas, Manuela Mollejo, Alberto Orfao, Luis Escribano
JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY
(2016)
Article
Dermatology
Adria Aterido, Antonio Julia, Carlos Ferrandiz, Lluis Puig, Eduardo Fonseca, Emilia Fernandez-Lopez, Esteban Dauden, Jose Luis Sanchez-Carazo, Jose Luis Lopez-Estebaranz, David Moreno-Ramirez, Francisco Vanaclocha, Enrique Herrera, Pablo de la Cueva, Nick Dand, Nuria Palau, Arnald Alonso, Maria Lopez-Lasanta, Rauel Tortosa, Andres Garcia-Montero, Laia Codo, Josep Lluis Gelpi, Jaume Bertranpetit, Devin Absher, Francesca Capon, Richard M. Myers, Jonathan N. Barker, Sara Marsal
JOURNAL OF INVESTIGATIVE DERMATOLOGY
(2016)
Letter
Oncology
I. Alvarez-Twose, P. Martinez-Barranco, J. Gotlib, A. Garcia-Montero, J. M. Morgado, M. Jara-Acevedo, J. D. Merker, F. J. Penalver, A. Matito, Y. Hou, L. Sanchez-Munoz, A. Mayado, M. Mollejo, L. Escribano, A. Orfao
Article
Rheumatology
Antonio Julia, Isidoro Gonzalez, Antonio Fernandez-Nebro, Francisco Blanco, Luis Rodriguez, Antonio Gonzalez, Juan D. Canete, Joan Maymo, Mercedes Alperi-Lopez, Alejandro Olive, Hector Corominas, Victor Martinez-Taboada, Alba Erra, Simon Sanchez-Fernandez, Arnald Alonso, Maria Lopez-Lasanta, Raul Tortosa, Laia Codo, Josep Lluis Gelpi, Andres C. Garcia-Montero, Jaume Bertranpetit, Devin Absher, S. Louis Bridges, Richard M. Myers, Jesus Tornero, Sara Marsal
Review
Oncology
Ivan Alvarez-Twose, Almudena Matito, Jose Mario Morgado, Laura Sanchez-Munoz, Maria Jara-Acevedo, Andres Garcia-Montero, Andrea Mayado, Carolina Caldas, Cristina Teodosio, Javier Ignacio Munoz-Gonzalez, Manuela Mollejo, Luis Escribano, Alberto Orfao
Article
Rheumatology
Antonio Julia, Francisco Blanco, Benjamin Fernandez-Gutierrez, Antonio Gonzalez, Juan D. Canete, Joan Maymo, Mercedes Alperi-Lopez, Alex Olive, Hector Corominas, Victor Martinez-Taboada, Isidoro Gonzalez-Alvaro, Antonio Fernandez-Nebro, Alba Erra, Simon Sanchez-Fernandez, Arnald Alonso, Maria Lopez-Lasanta, Raul Tortosa, Laia Codo, Josep Lluis Gelpi, Andres C. Garcia-Montero, Jaume Bertranpetit, Devin Absher, Richard M. Myers, Jesus Tornero, Sara Marsal
ARTHRITIS & RHEUMATOLOGY
(2016)
Article
Genetics & Heredity
Gabriella Galata, Andres C. Garcia-Montero, Thomas Kristensen, Ahmed A. Z. Dawoud, Javier Munoz-Gonzalez, Manja Meggendorfer, Paola Guglielmelli, Yvette Hoade, Ivan Alvarez-Twose, Christian Gieger, Konstantin Strauch, Luigi Ferrucci, Toshiko Tanaka, Stefania Bandinelli, Theresia M. Schnurr, Torsten Haferlach, Sigurd Broesby-Olsen, Hanne Vestergaard, Michael Boe Moller, Carsten Bindslev-Jensen, Alessandro M. Vannucchi, Alberto Orfao, Deepti Radia, Andreas Reiter, Andrew J. Chase, Nicholas C. P. Cross, William J. Tapper
Summary: Three intergenic SNPs associated with mastocytosis were identified, with one SNP showing association with age at presentation. These findings suggest that multiple germline variants predispose to KIT D816V positive mastocytosis and provide novel avenues for functional investigation.
AMERICAN JOURNAL OF HUMAN GENETICS
(2021)
Article
Hematology
Javier Munoz-Gonzalez, Ivan Alvarez-Twose, Maria Jara-Acevedo, Roberta Zanotti, Cecelia Perkins, Mohamad Jawhar, Wolfgang R. Sperr, Khalid Shoumariyeh, Juliana Schwaab, Georg Greiner, Ana Henriques, Carolina Caldas, Carlos Fernandez-Gimenez, Laura Sanchez-Munoz, Andrea Mayado, Alba Perez-Pons, Annette Schmitt-Graeff, Justus Duyster, Ilaria Tanasi, Francesco Olivieri, Elvira Mora-Castera, Irene Luna, Leonor Senent, Maria-Helena Banas, Amanda Nunez-Garcia, Manuel Jurado-Chacon, Guillermo Martin-Sanchez, Enrique Colado, Blanca Xicoy, Georgina Gener-Ricos, Jason Gotlib, Patrizia Bonadonna, Andreas Reiter, Peter Valent, Andres C. Garcia-Montero, Alberto Orfao
Summary: A risk stratification model for systemic mastocytosis was designed and validated, showing clear discrimination between low-risk and high-risk patients for progression-free survival (PFS) and overall survival (OS) outcomes. The model was based on unique combinations of independent prognostic factors and showed high accuracy in predicting PFS and OS in both non-advanced and advanced systemic mastocytosis.
LANCET HAEMATOLOGY
(2021)
Article
Hematology
Ana Henriques, Javier Munoz-Gonzalez, Laura Sanchez-Munoz, Almudena Matito, Lidia Torres-Rivera, Maria Jara-Acevedo, Carolina Caldas, Andrea Mayado, Alba Perez-Pons, Andres C. Garcia-Montero, Ivan Alvarez-Twose, Alberto Orfao
Summary: This study used highly sensitive flow cytometry to detect the presence of circulating tumor mast cells (CTMCs) in various types of systemic mastocytosis (SM) patients. CTMCs were found in higher counts in advanced SM and smoldering SM patients, but in lower counts in indolent SM and bone marrow mastocytosis patients. The presence of CTMCs was associated with adverse features, prognostic scores, and survival in SM patients.
Review
Oncology
Oscar Gonzalez-Lopez, Javier I. Munoz-Gonzalez, Alberto Orfao, Ivan Alvarez-Twose, Andres C. Garcia-Montero
Summary: Systemic mastocytosis (SM) is a rare clonal hematopoietic stem cell disease characterized by the expansion and accumulation of neoplastic mast cells carrying the activating KIT D816V mutation. Assessment of multilineage involvement of hematopoiesis by KIT D816V and additional mutations in genes known to be associated with the prognosis of SM can help identify high-risk patients who may benefit from closer monitoring and early treatment. Mutations in multiple genes, including ASXL1, CBL, DNMT3A, EZH2, NRAS, RUNX1, and SRSF2, have been found to be associated with poorer outcomes in SM patients.
Article
Allergy
David Gonzalez-de-Olano, Paula Navarro-Navarro, Javier I. Munoz-Gonzalez, Laura Sanchez-Munoz, Ana Henriques, Ana de-Andres-Martin, Dolores Peralta-Arjonilla, Andrea Mayado, Maria Jara-Acevedo, Andres C. Garcia-Montero, Alberto Orfao, Ivan Alvarez-Twose
Summary: This study aimed to determine the prevalence of hereditary alpha-tryptasemia (HAT) in healthy donors vs patients with different diagnostic subtypes of mast cell (MC) activation syndromes (MCAS) and mastocytosis, and its relationship with the clinical behavior of the disease.
Meeting Abstract
Hematology
A. Mayado, C. Teodosio, N. Dasilva-Freire, M. Jara-Acevedo, A. Garcia-Montero, I. Alvarez-Twose, L. Sanchez-Munoz, A. Matito, C. Caldas, J. Munoz-Gonzalez, A. Henriques, J. Sanchez-Gallego, L. Escribano, A. Orfao
Meeting Abstract
Hematology
C. Fernandez, C. Teodosio, S. Matarraz, A. Lopez, M. Jara-Acevedo, A. Mayado, I. Alvarez-Twose, L. Sanchez-Munoz, A. Garcia-Montero, M. L. Gutierrez, L. Escribano, A. Orfao