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丁香实验推荐阅读
Molecular Methods for Detection of Minimal Residual Disease Following Transplantation in Lymphoid and Plasma Cell Disorders

Relapse represents the main cause of treatment failure after stem cell transplantation (SCT). Thus, monitoring of minimal residual disease (MRD) in allografted patients allows an early detection of recurrence and a subsequent intervention prior to clinically detectable relaps ...

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Methods of Detection of Immune Reconstitution and T Regulatory Cells by Flow Cytometry

Allogeneic hematopoietic stem cell therapy (HSCT) remains one of the few curative treatments for high-risk hematological malignancies (high-risk leukemia, myelodysplastic syndromes, advanced myeloproliferative disorders, high-risk lymphomas, and multiple myelo ...

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Generation of Tissue Organoids by Compaction Reaggregation

Cellular reaggregation methods are commonly used to generate tissue organoids for use in biological studies. Using a modified method termed “compaction reaggregation,” it is possible to establish reaggregates of reproducible size from defined input cell numbers with ease and wit ...

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Generation of Mouse Embryos with Small Hairpin RNA-Mediated Knockdown of Gene Expression

We are using knockdown of gene expression in mouse embryos by constitutive expression of small hairpin (sh)RNAs as a means of observing loss-of-function phenotypes more rapidly than gene targeting. Plasmid constructs that direct shRNA expression via an RNA pol III promoter are introduc ...

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Identifying Essential Genes in Mouse Development via an ENU-Based Forward Genetic Approach

The completion of the human and mouse genome projects at the beginning of the past decade represented a very important step forward in our pursuit of a comprehensive understanding of the genetic control of mammalian development. Nevertheless, genetic analyses of mutant phenotypes are st ...

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Real-Time PCR Quantification of Gene Expression in Embryonic Mouse Tissue

The Gbx family of transcription factors consists of two closely related proteins GBX1 and GBX2. A defining feature of the GBX family is a highly conserved 60 amino acid DNA-binding domain, which differs by just two amino acids. Gbx1 and Gbx2 are co-expressed in several areas of the developing central ...

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EMAGE: Electronic Mouse Atlas of Gene Expression

The EMAGE (Electronic Mouse Atlas of Gene Expression) database (http://www.emouseatlas.org/emage) allows users to perform on-line queries of mouse developmental gene expression. EMAGE data are represented spatially using a framework of 3D mouse embryo models, thus allowing uni ...

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Laser Capture Microdissection of Embryonic Cells and Preparation of RNA for Microarray Assays

In order to compare the global gene expression profiles of different embryonic cell types, it is first necessary to isolate the specific cells of interest. The purpose of this chapter is to provide a step-by-step protocol to perform laser capture microdissection (LCM) on embryo samples and obt ...

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Detection and Monitoring of MicroRNA Expression in Developing Mouse Brain and Fixed Brain Cryosections

MicroRNAs (miRNAs) are 20−25 nucleotide long, noncoding, and single-strand RNAs that have been found in almost all organisms and shown to exert essential roles by regulating the stability and translation of target mRNAs. In mammals most miRNAs show tissue specific and developmentally re ...

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Two-Color In Situ Hybridization of Whole-Mount Mouse Embryos

RNA in situ hybridization is a powerful technique used to identify the spatial localization of a specific RNA in a tissue section or whole tissue. In this protocol, we describe a reliable method for two-color in situ hybridization that can be used to accurately assess the expression of multiple gen ...

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Essentials of Recombinase-Based Genetic Fate Mapping in Mice

Fate maps, by defining the relationship between embryonic tissue organization and postnatal tissue structure, are one of the most important tools on hand to developmental biologists. In the past, generating such maps in mice was hindered by their in utero development limiting the physical ...

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Genetic Cell Ablation

Targeted cell ablation has proven to be a valuable approach to study in vivo cell functions during organogenesis, tissue homeostasis, and regeneration. Over the last two decades, various approaches have been developed to refine the control of cell ablation. In this review, we give an overview of ...

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Live Imaging Mouse Embryonic Development: Seeing Is Believing and Revealing

The use of genetically encoded fluorescent proteins has revolutionized the fields of cell and developmental biology and redefined our understanding of the dynamic morphogenetic processes that work to shape the embryo. Fluorescent proteins are routinely used as vital reporters to ...

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Using the Textpresso Site-Specific Recombinases Web Server to Identify Cre Expressing Mouse Strains and Floxed Alleles

Effective tools for searching the biomedical literature are essential for identifying reagents or mouse strains as well as for effective experimental design and informed interpretation of experimental results. We have built the Textpresso Site Specific Recombinases (Textp ...

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Analyzing Gene Function in Whole Mouse Embryo and Fetal Organ In Vitro

A well-established experimental paradigm to analyze gene function in development is to elucidate the impact of gain and loss of gene activity on cell differentiation, tissue modelling, organogenesis, and morphogenesis. This chapter describes the experimental protocols to study ...

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The Latest Improvements in the Mouse Sperm Preservation

Sperm preservation is an important technique for maintaining valuable genetic resources in biomedical research and wildlife. In the mouse, the sperm cryopreservation method has been established and adopted by large-scale sperm preservation projects in cryobanks. Recently, a n ...

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Collection and Preparation of Rodent Embryonic Samples for Transcriptome Study

The need for large-scale collection of rodent embryos and individual embryonic tissues for genomic and proteomic studies requires modification of traditional practices of embryo necropsy. The sample intended for transcriptome study should be rapidly dissected and stabilized ...

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In Situ Hybridization Methods for Mouse Whole Mounts and Tissue Sections with and Without Additional -Galactosidase Staining

In situ hybridization is a powerful method for detecting endogenous mRNA sequences in morphologically preserved samples. We provide in situ hybridization methods, which are specifically optimized for mouse embryonic samples as whole mounts and section tissues. Additionally, β ...

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Microscopic Computed Tomography-Based Virtual Histology of Embryos

Advances in imaging technologies and computational capabilities have made possible novel methods for phenotypic assessments and visualization of detailed anatomical structures of whole embryos. We recently reported a rapid and inexpensive technique for achieving high-r ...

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Detection of Cells Programmed to Die in Mouse Embryos

Programmed Cell Death (PCD) is a broad term used to describe a series of events that culminate in the death of specific cells. In the embryo it occurs at predictable stages and tissues. During mouse development, PCD is a mechanism to preserve the homeostasis of the growing organism, and also is needed for t ...

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