Acid Fast Staining is used to identify acid fast organisms such mycobacteria in paraffin-embedded tissue section. Acid fast bacteria have a high content of mycolic acids in their cell walls. Acid fast bacteria will be red, while nonacid fast bacteria will stain blue/green with a counterstain.

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Alcian blue stains acid mucosubstances and acetic mucins in paraffin-embedded tissue section. Excessive amounts of non-sulfated acidic mucosubstances are seen in mesotheliomas, certain amounts occur normally in blood vessel walls but increase in early lesions of atherosclerosis. Results: acid mucins/mucosubstances: blue; Nuclei (using nuclear fast red) redish pink.

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To obtain a good monoclonal antibody it is critical to have right screen method(s). We can help with your antibody selection and characterization by: 1. Design appropriate tissue panels or TMAs for your antibody initial screening; 2. Determine the tissue/cell specificity and cross-reactivity of the selected clones; 3. Establish optimal protocols for your final clones for use in IHC; 4. Compare the IHC staining of your antibodies with that of existing ones to the same target(s); 5. Provide reports or documents for the above characterization.

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Taking picture of a selected area under bright field microscope by a pathologist

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Cell Block Preparation including cell fixation, palleting, pellet processing and embedding

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For many new drugs, it is critical to have appropriate companion diagnostic tests to aid in selecting or excluding patients for the treatment, based on expression patterns of certain biomarkers in the diseased tissues that help predict likely response or severe toxicity. With our tissue bank and technology platforms, we can provide: 1. Validation of your companion biomarkers in hundreds or thousands of cases of a relevant disease tissue samples; 2. Screening and develop control panels or TMAs for standardization of the companion tests.

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Preparation of cytospin slide from cell suspension including fixation.

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Bone decalcification with EDTA based solution to prepare bone sample suitable for sectioning and various staining

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Probe labeling using in vitro transcription or other methods

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Double immunofluorescent IHC staining procedures from dewaxing to mouting for a single tissue, cell or TMA section. Detection reagents are free for mouse and rabbit primary antibodies.

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FISH with commercial kit for RNA and DNA target on tissue sections

200

Taking picture of a selected area under fluorescence microscope by a pathologist

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H&E staining on paraffin-embedded tissue section

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Double IHC staining procedures from dewaxing to mouting for a single tissue section or TMA section. Detection reagents are free for mouse and rabbit primary antibodies.

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Standard DAB based IHC staining procedures from dewaxing to mouting for a single tissue section or large size tissue array section. Detection reagents are free for mouse and rabbit primary antibodies.

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Standard DAB based IHC staining procedures from dewaxing to mouting for a single tissue section or medium size tissue array section. Detection reagents are free for mouse and rabbit primary antibodies.

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Standard DAB based IHC staining procedures from dewaxing to mouting for a single tissue, cell or small TMA section. Detection reagents are free for mouse and rabbit primary antibodies.

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IHC procedures including tests for primary antibody concentrations and controls on our universal tissue arrays to set up an optimal IHC protocol. Detection reagents and universal tissue arrays are free for mouse and rabbit primary antibodies.

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IHC staining procedures with alkaline phosphatase and its substate from dewaxing to mouting for a single tissue, cell or TMA section. Detection reagents are free for mouse and rabbit primary antibodies.

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Perls' Prussian Blue Iron Staining demonstrates ferric iron paraffin-embedded tissue section. Results: iron - blue; nuclei - red; background - pink.

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Standard ISH procedures for DNA target from dewaxing to mouting for a single tissue section or small tissue array section. Detection reagents are free for digoxigenin labeled probes.

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Standard ISH procedures for RNA target from dewaxing to mouting for a single tissue section or small tissue array section. Detection reagents are free for digoxigenin labeled probes.

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ISH procedures with RNAscope� Kit for detection of RNA target from dewaxing to mouting for a single tissue section or small tissue array section.

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ISH procedures including at least 3 tests for probe and enzyme concentrations on our universal tissue arrays. Detection reagents and universal tissue arrays are free for digoxigenin labeled probes.

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PAS staining on paraffin-embedded tissue section. This method stains polysaccharides such as glycogen, and mucosubstances such as glycoproteins, glycolipids and mucins in tissues. Glycogen, neutral mucosubstances, basement membranes, collagen fibers, glycolipids and phospholipids are demonstrated as pink to red to purple color.

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With our comprehensive tissue bank, we can organize tissue samples relevant to a disease process, a pathological feature or a group of pathological conditions into pathology panels or models. IHC or ISH analysis with appropriate antibodies or RNA probes of these panels or models (in either single tissue sections or TMAs) allows easy observation, comparison and correlation of target expression patterns and the disease process under investigation. All these will form an effective approach to verify disease associated targets and eliminate non-relevant targets even at very early stage of drug discovery. 1. Inflammatory models: We can provide tissue samples covering a wide spectrum of inflammatory process, from acute reaction to chronic inflammation either related to multiple organs or related to a specific organ, such as COPD lesions, various types of hepatitis. 2. Models for diseases of immunity: We can organize tissue samples for autoimmune disease, such as Graves diseases, psoriasis for our customers. 3. Oncology models: We can organize samples into different panels for validation of targets that may be associated with a). evolution from premalignant to malignant; b). progression from low grade to high grade; c). differences between normal and tumor; d). differences between primary and metastatic tumors; e). differences between good treatment response and poor response; f). occurrence of apoptosis; g). development of angiogenesis.

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Description and comments on any tissue (human or animal) section-based staining or study results by a ABP certified or non-ABP certified pathologist

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With commercial image analysis software, we can provide accurate quantitative analysis for nuclear, membrane and cytoplasmic markers identified by IHC, IF, ISH or special staining on cell smears/spins and tissue sections. 1. Cell number counting - positive cells per section, per TMA core or per high power view; 2. H-Score analysis � Software will identify and record negative (- or 0), weak (+ or 1), moderate (++ or 2) and strong (+++ or 3) staining intensity, and positive rate (%) of each intensity category for the target cells across the section or view under analysis. H-Score is then calculated by the formula: (3 x percentage of strongly stained target cells) + (2 x percentage of moderately stained target cells) + (1 x percentage of weakly stained target cells), giving a range of 0 to 300. 3. Positive IHC (protein), ISH (RNA) or special staining area vs negative area across the whole section or region of interest (ROI). Our software can identify different color, texture and contextual features, and segment them into appropriate areas (such as normal, tumor, necrosis, fibrosis or stromal areas) that can be analyzed (into percentage) and compared.

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DNA and RNA probe design for ISH

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Cutting single tissue section on charged slide at 4 um thickness

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Cutting section with special requirement

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Cutting large size array section on charged slide at 4um thickness

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Cutting medium size array section on charged slide at 4um thickness

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Cutting small size array section on charged slide at 4um thickness

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Direct scoring by eyes under a microscope is the most preferred and least expensive method for IHC, IF and ISH staining analysis. We provide the following semi-quantitative analyses for nuclear, membrane and cytoplasmic markers on cell smears/spins and tissue sections: 1. Basic analysis � including negative (- or 0), weak (+ or 1), moderate (++ or 2) and strong (+++ or 3) for average staining intensity, and positive rate (%) of the target cells across the section under analysis. 2. Semi-quantitative H-Score analysis - This is obtained by the formula: (3 x percentage of strongly stained target cells) + (2 x percentage of moderately stained target cells) + (1 x percentage of weakly stained target cells), giving a range of 0 to 300. 3. Detailed semi-quantitative analysis � Detailed staining intensity results are recorded as: �0� is negative. �0.5� is borderline staining with no significance. �1� is weak staining. �1.5� is weak staining with foci of moderate staining. �2� is moderate staining. �2.5� is moderate staining with foci of strong staining. �3� is homogeneous strong staining. �3.5� is very strong and homogeneous staining with no significant background. �4� is over staining usually with background staining. Any figure in between can be described towards the description of the higher end, for example, 2.7 can be considered as strong staining. The average positive rate (%) of the target cells across each section is also recorded.

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Silver staining is used to identify fungi in paraffin-embedded tissue section. The mucopolysaccharide components of the fungal cell wall are oxidized to release aldehyde groups. The aldehyde groups then react with the silver nitrate, reducing it to a metallic silver, rendering them visible. Results: fungi - black; background - green.

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For all new therapeutic antibodies, it is required by regulatory agencies to assess their safety by examining and identifying their potential cross reactivity (TCR) on a panel of 33 normal tissue types from at least three individuals. Following FDA recommendations, we can provide IHC based-TCR examination on: 1. Single tissue sections of 33 normal tissue types on three or more individuals; 2. Low cost TMA of 33 normal tissue types on one individual (MNO341)l; 3. TMA of 33 normal tissue types in duplicates (MNO661); 4. TMA of 33 normal tissue types of three individuals (MNO961 or MNO1021); 5. *Frozen TMA of 33 normal tissue types of one individual (total three sets of TMA from three individuals). �*� � under development.

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Tissue processing procedures including trimming, cassettibg, dehydrating, paraffin-embedding

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Re-embedding customer's block

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We can provide FFPE single tissue blocks or TMA blocks of any format from common experimental animals, such as mouse (all the common strains), rat, rabbit, hamster, guinea pig, dog, monkey. With our dedicated tissue processing center, we can fix, process and make these animal tissues into paraffin blocks according to customers� needs. With our tissue bank and hospital links, we can also organize a wide range of normal and diseased human FFPE tissues (from a few cases to over 1000 cases) for our customers following appropriate technical, ethic guidelines and consents.

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We can repair any damage to a TMA paraffin block, such as craks, uneven surface, and shap alteration. We can also replace/refill any missing or unwanted core with an appropirate donor block

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Arraying/coring from a donor block to a recipient block with special conditions, such as small biopsy samples, thin samples or hard to define samples

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Arraying/coring from donor block to recipient block, per 4.5mm core

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Arraying/coring from donor block to recipient block, per 2mm or 2.5mm core

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Arraying/coring from donor block to recipient block, per 1mm or 1.5mm core

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Transfer a marked spot from a slide to a donor block

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Select and mark a spot on a stained slide by our pathologist

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Procedures needed for initiation of a TMA construction including machine alignment, calibration, and material/design verification.

100

Trichrome staining is used to differentiate between collagen and smooth muscle in tumors, and is also used to outline the increase of collagen in diseases such as cirrhosis. Results: nuclei - bloack; cytoplasm, muscle, erythrocytes - red; colleagen - blue.

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Bright field image scanning of a whole slide with a single tissue section with Aperio slide scanner. Price starts from $48 per routine slide and may vary for magnifications and slide numbers.

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Bright field image scanning of a whole TMA slide with Aperio slide scanner. Price starts from $80 per TMA slide and may vary for magnifications and slide numbers.

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