The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

Leica Biosystems Accessory

Leica Biosystems Accessory

Leica Biosystems Accessory

Leica Biosystems Accessory

Leica Biosystems Accessory

The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

The instrument has been designed so that it is safe to use by the operator as well as for processing speci-mens—provided that it is operated according to these Instructions for Use. The Leica TP1020 is a modular automated tissue processor designed for the following laboratory applications: • Fixation • Dehydration • Paraffin infiltration of histological tissue specimens. The Leica TP1020 must be operated exclusively with the reagents listed in the IFU

Leica Biosystems Accessory

Leica Biosystems Accessory

HistoCore SPECTRA Accessory

The HistoCore PEGASUS is an automated tissue processor specifically designed for fixation, dehydration, infiltration with intermedium and paraffin infiltration of human tissue specimens used for histological medical diagnosis by a pathologist, e.g. for cancer diagnosis. The HistoCore PEGASUS is designed for in vitro diagnostic applications.

The HistoCore PEGASUS Plus is an automated tissue processor specifically designed for fixation, dehydration, infiltration with intermedium and paraffin infiltration of human tissue specimens used for histological medical diagnosis by a pathologist, e.g. for cancer diagnosis. The HistoCore PEGASUS Plus is designed for in vitro diagnostic applications.

HistoCore PEARL Accessory

The HistoCore PEARL is designed to enable tissue processing, performing all steps from fixation to wax infiltration. It is normally processed in sequential mode, which means that the instrument is strictly using one bottle after the other. The HistoCore PEARL is a tissue processor for the following laboratory applications: Fixation Dehydration Infiltration with intermedium Paraffin infiltration of histological tissue specimens. The HistoCore PEARL must be operated exclusively with the reagents listed in the section of Compatible reagents

The ASP6025 Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 can be used as medical IVD product. The improved features of the ASP6025 are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The ASP6025 Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 can be used as medical IVD product. The improved features of the ASP6025 are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

HistoCore PEARL Accessory

The ASP6025 Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 can be used as medical IVD product. The improved features of the ASP6025 are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The ASP6025 Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 can be used as medical IVD product. The improved features of the ASP6025 are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The ASP6025 S Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 S can be used as medical IVD product. The improved features of the ASP6025 S are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The ASP6025 S Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 S can be used as medical IVD product. The improved features of the ASP6025 S are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The ASP6025 S Tissue Processor is a modular tissue processor for the following laboratory applications: • Fixation • Dehydration • Infiltration with intermedium • Paraffin infiltration of histological tissue specimens. The ASP6025 S can be used as medical IVD product. The improved features of the ASP6025 S are the combination of: • Reduced processing time • Easier reagent exchange • Monitoring the ethanol quality by density measurement as well as the • Ability to replace xylene, a substance that is hazardous (including to the health), with special process protocols. The instrument has been designed so that it is safe to use by the operator as well as for processing specimens. However, the prerequisite for this is that it is operated according to these Instructions for Use.

The Leica Small Linear Stainer ST4020 is used for the automated preparation of routine histo¬logical and cytological stains. It is i ntended for use in pathology laboratories to assist with the following tasks: • Staining thin tissue sections or cytological specimens on slides. Frozen sections can also be stained. • The instrument may be operated only ccording to the instructions contained in this manual.

Leica Biosystems Accessory

Leica Biosystems Accessory

The Leica Small Linear Stainer ST4020 is used for the automated preparation of routine histo¬logical and cytological stains. It is i ntended for use in pathology laboratories to assist with the following tasks: • Staining thin tissue sections or cytological specimens on slides. Frozen sections can also be stained. • The instrument may be operated only ccording to the instructions contained in this manual.

HistoCore SPECTRA Accessory

The HistoCore SPECTRA ST is an in-vitro diagnostics instrument. The HistoCore SPECTRA ST is a stainer for laboratory applications and intended for the preparation of histological and cytological specimens.

The HistoCore SPECTRA CV is a fully automated coverslipper that is used to apply mounting media between the slide and coverglass. Then a coverglass is applied in order to preserve conserve the specimen and to create a uniform visual surface for microscopic research on histological and cytological tissue samples for medical diagnostics (e.g. cancer diagnostics). This instrument may be operated only in conjunction with the coverglasses and mounting media approved by Leica.

The Stainer Leica ST5020 is an automated stain-er for the preparation of histological and cyto-logical routine stainings. It is designed for use in pathology laboratories, and only for performing the following tasks: • Staining of thin sections of tissue specimens or of cytological samples, attached to micro-scope slides • The instrument may be operated only ac¬cording to the instructions contained in this manual.

The Stainer Leica ST5020 is an automated stain-er for the preparation of histological and cyto-logical routine stainings. It is designed for use in pathology laboratories, and only for performing the following tasks: • Staining of thin sections of tissue specimens or of cytological samples, attached to micro-scope slides • The instrument may be operated only ac¬cording to the instructions contained in this manual.

REA747 recognizes the human CD45 antigen, a member of the protein tyrosine phosphatase (PTP) family, also known as the leukocyte common antigen (LCA). The CD45 molecule a type I transmembrane protein which is required for T and B cell activation and involved in cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45 is expressed in different isoforms (CD45RA, CD45RB, CD45RC, CD45RAB, CD45RAC, CD45RBC, CD45RO, CD45R (ABC)) depending on the differentiation status of the cell. The CD45 antibody recognizes a common epitope of all CD45 isoforms. Additional information: Clone REA747 displays negligible binding to Fc receptors.

REA747 recognizes the human CD45 antigen, a member of the protein tyrosine phosphatase (PTP) family, also known as the leukocyte common antigen (LCA). The CD45 molecule a type I transmembrane protein which is required for T and B cell activation and involved in cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45 is expressed in different isoforms (CD45RA, CD45RB, CD45RC, CD45RAB, CD45RAC, CD45RBC, CD45RO, CD45R (ABC)) depending on the differentiation status of the cell. The CD45 antibody recognizes a common epitope of all CD45 isoforms. Additional information: Clone REA747 displays negligible binding to Fc receptors.

AC136 recognizes the human CD34, a class III epitope of the CD34 antigen. This epitope is different than the one recognized by the clone used in the CD34 MicroBead Kits. The CD34 antigen is a single-chain transmembrane glycoprotein, expressed on human hematopoietic stem and progenitor cells, endothelial progenitor cells, vascular endothelial cells, embryonic fibroblasts, and some cells in fetal and adult nervous tissue. The antigen is absent on fully differentiated hematopoietic cells, such as normal peripheral blood lymphocytes, monocytes, granulocytes, erythrocytes, and platelets. Clone AC136 has a similar specificity as the CD34 monoclonal antibody clone 8G12 (HPCA-2). CD34 antibodies can be used for studies of hematopoiesis and nonhematopoietic stem cells, phenotyping of hematopoietic stem cells, and studies on phenotyping of hematologic malignancies, endothelial cells, and endothelial progenitor cells (EPCs).

T6D11 recognizes the human CD45RA antigen which is expressed on naive CD4+ and CD8+ T cells as well as on CD8+ effector T cells. CD45RA is also present on subsets of B and NK cells as well as on plasmacytoid dendritic cells. The CD45RA antibody recognizes the 220 kDa isoform of the leukocyte common antigen (LCA), a transmembrane tyrosine phosphatase.

145/15 recognizes the human CD62L antigen, a 74 kDa glycoprotein and a member of the selectin family of cell surface molecules. CD62L is also known as L-selectin, LECAM-1, or LAM-1 and binds a series of glycoproteins including CD34, GlyCAM-1, and MAdCAM-1. CD62L is important for homing of naive lymphocytes via endothelial venules to peripheral lymph nodes and Peyer’s patches. The CD62L antigen also contributes to the recruitment of leukocytes from the blood to areas of inflammation. Most hematopoietic cells express CD62L, including many peripheral blood B cells, T cells, monocytes, granulocytes, and some myeloid cells from bone marrow, and thymocytes. CD62L is continuously endoproteolytically cleaved from the cell surface neutrophils and lymphocytes (shedding). Proteolysis is accelerated, for example, after antigenic activation of T cells. Always use fresh material for immunofluorescent staining of CD62L+ cells. For optimal results, the cells should not be older than 8–12 hours. Keep cells continuously cold. CD62L-expression may be rapidly lost due to shedding.

145/15 recognizes the human CD62L antigen, a 74 kDa glycoprotein and a member of the selectin family of cell surface molecules. CD62L is also known as L-selectin, LECAM-1, or LAM-1 and binds a series of glycoproteins including CD34, GlyCAM-1, and MAdCAM-1. CD62L is important for homing of naive lymphocytes via endothelial venules to peripheral lymph nodes and Peyer’s patches. The CD62L antigen also contributes to the recruitment of leukocytes from the blood to areas of inflammation. Most hematopoietic cells express CD62L, including many peripheral blood B cells, T cells, monocytes, granulocytes, and some myeloid cells from bone marrow, and thymocytes. CD62L is continuously endoproteolytically cleaved from the cell surface neutrophils and lymphocytes (shedding). Proteolysis is accelerated, for example, after antigenic activation of T cells. Always use fresh material for immunofluorescent staining of CD62L+ cells. For optimal results, the cells should not be older than 8–12 hours. Keep cells continuously cold. CD62L-expression may be rapidly lost due to shedding.

REA546 recognizes the human CD197 antigen, which is also known as CCR7. CD197 is a chemokine receptor with a C-C motif that mediates homing of T cells to secondary lymphoid organs via high endothelial venules (HEV). For example, naive T cells migrate very efficiently through lymph nodes using the adhesion molecules L-selectin, CD62L, and CCR7. Ligands for CCR7, e.g. CCL19, are expressed by the HEV of secondary lymphoid organs, by parenchymal cells within T cell zones of lymph nodes, and by endothelial cells at the openings of lymphatic vessels within peripheral tissues. Expression of CCR7 and the CD45RA isoform distinguishes three subsets of T cells: naive T cells (CCR7+CD45RA+), central memory T cells (CCR7+ CD45RA-), and effector memory T cells (CCR7- CD45RA-). Additional information: Clone REA546 displays negligible binding to Fc receptors.

REA623 recognizes the human CD4 antigen, a 55 kDa single-pass type I membrane protein, also known as T4/Leu-3. CD4 is highly expressed on T helper cells and at a lower level on monocytes and dendritic cells. It is involved in the recognition of MHC class ll/peptide complexes by the TCR heterodimers and is the receptor for the human immunodeficiency virus (HIV). Additional information: Clone REA623 displays negligible binding to Fc receptors.

11F2 recognizes the human gamma/delta T cell receptor (TCR). The T cell receptor is a heterodimeric glycoprotein associated with the CD3 antigen. The TCR consists of a alpha and a beta chain (TCRa/b) or a gamma and a delta chain (TCRg/d). Clone 11F2 reacts with a framework epitope of the gamma/delta T cell–receptor. The gamma and delta TCR chains are composed of constant and variable regions, each encoded by distinct gene segments. The gamma chain forms either disulfide-linked or non-disulfide-linked heterodimers with the delta-subunit. The gamma/delta T cell–receptor is present on a subset of T lymphocytes in peripheral blood, intestinal epithelium, lymph node, thymus, and spleen. TCR gamma/delta is involved in the recognition of certain bacterial, self-CD1 molecule, and tumor antigens bound to MHC class I. Gamma/delta T cells are mainly CD4 negative and CD8 negative. T cells expressing the gamma/delta TCR have been shown to play a role in oral tolerance, innate immune response for some tumor cells, and autoimmune disease. Antigen presentation by gamma/delta T cells has been reported.

REA1047 recognizes the human CD45FtA antigen, which is expressed on naive CD4+ and CD8+ T cells as well as on CD8+ effector T cells. CD45RA is also present on subsets of B and NK cells as well as on plasmacytoid dendritic cells. The CD45FtA antibody recognizes the 220 kDa isoform of the leukocyte common antigen (LCA), a transmembrane tyrosine Phosphatase. Additional information: Clone REA1047 displays negligible binding to Fc receptors.

Aged human EDTA-treated whole blood samples (6 days old) were stained with 7-AAD Staining Solution using a lyse-wash procedure and analyzed by flow cytometry using the MACSQuant® Analyzer. For analysis, white blood cells were gated based on forward and side scatter properties and after exclusion of cell debris and red blood cells based on scatter signals. Dead and apoptotic cells were identified by 7-AAD fluorescence.