diff --git a/luminex/src/org/labkey/luminex/LuminexUploadWizardAction.java b/luminex/src/org/labkey/luminex/LuminexUploadWizardAction.java
index a95fe9f5a..d5b09d44c 100644
--- a/luminex/src/org/labkey/luminex/LuminexUploadWizardAction.java
+++ b/luminex/src/org/labkey/luminex/LuminexUploadWizardAction.java
@@ -444,8 +444,10 @@ public void writeSameCheckboxCell(RenderContext ctx, HtmlWriter out)
                     ).appendTo(out);
 
                     StringBuilder onchange = new StringBuilder("b = this.checked;");
-                    for (DisplayColumn col : getColumns())
+                    // Index starts at 1 -- always leave the first column visible (Issue 53620)
+                    for (int i = 1; i < getColumns().size(); i++)
                     {
+                        DisplayColumn col = getColumns().get(i);
                         onchange.append("document.getElementsByName('").append(col.getFormFieldName(ctx)).append("')[0].style.display = b ? 'none' : 'block';\n");
                     }
                     onchange.append("if (b) { ").append(groupName).append("Updated(); }");
diff --git a/luminex/src/org/labkey/luminex/query/NegativeBeadDisplayColumnGroup.java b/luminex/src/org/labkey/luminex/query/NegativeBeadDisplayColumnGroup.java
index d6665d695..e10842759 100644
--- a/luminex/src/org/labkey/luminex/query/NegativeBeadDisplayColumnGroup.java
+++ b/luminex/src/org/labkey/luminex/query/NegativeBeadDisplayColumnGroup.java
@@ -51,7 +51,10 @@ public void writeSameCheckboxCell(RenderContext ctx, HtmlWriter out)
                 InputBuilder.checkbox().name(id).id(id).appendTo(out);
                 StringBuilder onChange = new StringBuilder("b = this.checked;\n");
 
-                getColumns().forEach(col -> {
+                // Index starts at 1 -- always leave the first column visible (Issue 53620)
+                for (int i = 1; i < getColumns().size(); i++)
+                {
+                    DisplayColumn col = getColumns().get(i);
                     if (col.getColumnInfo() != null)
                     {
                         onChange.append("s = document.getElementsByName('")
@@ -61,7 +64,7 @@ public void writeSameCheckboxCell(RenderContext ctx, HtmlWriter out)
                             .append(col.getFormFieldName(ctx))
                             .append("')[0].style.display = b || s == 0 ? 'none' : 'block';\n");
                     }
-                });
+                }
 
                 onChange.append(" if (b) { ")
                     .append(inputName)
diff --git a/luminex/test/src/org/labkey/test/tests/luminex/LuminexRTransformTest.java b/luminex/test/src/org/labkey/test/tests/luminex/LuminexRTransformTest.java
index c2489e0f2..f4c574adf 100644
--- a/luminex/test/src/org/labkey/test/tests/luminex/LuminexRTransformTest.java
+++ b/luminex/test/src/org/labkey/test/tests/luminex/LuminexRTransformTest.java
@@ -127,6 +127,7 @@ public void uploadRun()
         setFormElement(Locator.name("StndCurveFitInput"), "FI");
         setFormElement(Locator.name("UnkCurveFitInput"), "FI-Bkgd-Neg");
         setFormElement(Locator.name("__primaryFile__"), TEST_ASSAY_LUM_FILE4);
+        scrollIntoView(Locator.lkButton("Next"), true);
         clickButton("Next", defaultWaitForPage * 2);
 
         // make sure the Standard checkboxes are checked
@@ -140,12 +141,11 @@ public void uploadRun()
         setFormElement(Locator.xpath("//input[@type='text' and contains(@name, '_LotNumber')][1]"), TEST_ANALYTE_LOT_NUMBER);
         // set negative control and negative bead values
         checkCheckbox(Locator.name("_analyte_" + ANALYTE3 + "_NegativeControl"));
+        checkCheckbox(Locator.name("_analyte_" + ANALYTE1 + "_NegativeBeadCheckBox")); // Issue 53620: "Same" checkbox for subtract negative bead
         selectOptionByText(Locator.name("_analyte_" + ANALYTE1 + "_NegativeBead"), ANALYTE3);
-        selectOptionByText(Locator.name("_analyte_" + ANALYTE2 + "_NegativeBead"), ANALYTE3);
         // switch to using MyNegative bead for subtraction
         checkCheckbox(Locator.name("_analyte_" + ANALYTE4 + "_NegativeControl"));
         selectOptionByText(Locator.name("_analyte_" + ANALYTE1 + "_NegativeBead"), ANALYTE4);
-        selectOptionByText(Locator.name("_analyte_" + ANALYTE2 + "_NegativeBead"), ANALYTE4);
         clickButton("Save and Finish");
     }
 
diff --git a/luminex/test/src/org/labkey/test/tests/luminex/LuminexTest.java b/luminex/test/src/org/labkey/test/tests/luminex/LuminexTest.java
index b61ac90e9..165c17756 100644
--- a/luminex/test/src/org/labkey/test/tests/luminex/LuminexTest.java
+++ b/luminex/test/src/org/labkey/test/tests/luminex/LuminexTest.java
@@ -662,6 +662,7 @@ public void selectPositivityFile(File file, boolean expectDuplicateFile)
         setFormElement(Locator.name("__primaryFile__"), file);
         if (expectDuplicateFile)
             waitForText("A file with name '" + file.getName() + "' already exists");
+        scrollIntoView(Locator.lkButton("Next"), true);
         clickButton("Next");
     }